Khattak, Shahryar; Tanaka, Elly M
Transgenic animals have been indispensable in elucidating and deciphering mechanisms underlying various biological phenomena. In regeneration, transgenic animals expressing fluorescent protein genes have been crucial for identifying the source cells for regeneration and the mechanism of blastema formation. Animals are usually generated by manipulating their genome using various techniques at/in one cell embryo/fertilized egg stage. Here, we describe the generation of germline transgenic axolotls (Ambystoma mexicanum) using the I-SceI meganuclease and Tol2 transposase.
Hoffman, Robert L.; Larson, Gary L.; Brokes, Brendan J.
Ambystoma gracile (Baird) and Ambystoma macrodactylum Baird are common salamander species occupying key trophic positions in mountain ponds and lakes of Mount Rainier National Park (MORA). Based on amphibian surveys conducted in ponds and lakes in the northeast quadrant of MORA during the summers of 1993a??1995, 20 sites were resampled in 1996 to document and evaluate distributions of the two species relative to site habitat characteristics. Distributions of larvae were assessed either by nearshore snorkel or visual encounter surveys of each study site. Twelve environmental variables representing pond and lake physical characteristics, water quality, and nutrient concentrations were measured. The occurrences at each site of three major habitat components (i.e., pond bottom firmness, coarse woody debris, and emergent/aquatic vegetation) were qualitatively estimated. Allotopic distributions of larval populations of both species were related to site elevation, maximum depth, organic content of bottom substrates, and surface area. Ambystoma gracile sites were larger, deeper, lower in elevation, had flocculent sediment higher in organic content, abundant coarse woody debris, and limited emergent/aquatic vegetation relative to A. macrodactylum sites. Ambystoma macrodactylum sites were smaller, shallower, higher in elevation, had firm sediments low in organic content, and had a greater occurrence of emergent/aquatic vegetation than did A. gracile sites. Two sites supported syntopic populations and exhibited many of the habitat characteristics observed at sites inhabited by each species. The distribution of each species in MORA was related to the interconnection between habitat characteristics of ponds and lakes and species life-history patterns and possibly interspecific interactions.
Voss S Randal
Full Text Available Abstract Background Because amphibians arise from a branch of the vertebrate evolutionary tree that is juxtaposed between fishes and amniotes, they provide important comparative perspective for reconstructing character changes that have occurred during vertebrate evolution. Here, we report the first comparative study of vertebrate genome structure that includes a representative amphibian. We used 491 transcribed sequences from a salamander (Ambystoma genetic map and whole genome assemblies for human, mouse, rat, dog, chicken, zebrafish, and the freshwater pufferfish Tetraodon nigroviridis to compare gene orders and rearrangement rates. Results Ambystoma has experienced a rate of genome rearrangement that is substantially lower than mammalian species but similar to that of chicken and fish. Overall, we found greater conservation of genome structure between Ambystoma and tetrapod vertebrates, nevertheless, 57% of Ambystoma-fish orthologs are found in conserved syntenies of four or more genes. Comparisons between Ambystoma and amniotes reveal extensive conservation of segmental homology for 57% of the presumptive Ambystoma-amniote orthologs. Conclusion Our analyses suggest relatively constant interchromosomal rearrangement rates from the euteleost ancestor to the origin of mammals and illustrate the utility of amphibian mapping data in establishing ancestral amniote and tetrapod gene orders. Comparisons between Ambystoma and amniotes reveal some of the key events that have structured the human genome since diversification of the ancestral amniote lineage.
Calfee, R.D.; Bridges, C.M.; Little, E.E.
Increased ultraviolet-B (UV-B) radiation reaching the Earth's surface has been implicated in amphibian declines. Recent studies have shown that many amphibian species have differences in sensitivity depending on developmental stage. Embryos and larvae of Ambystoma maculatum (Spotted Salamander) and larvae of Ambystoma talpoideum (Mole Salamander) were exposed to five simulated UV-B treatments in controlled laboratory experiments to determine the relative sensitivity of different lifestages. Hatching success of the embryos exceeded 95% in all treatments; however, the larvae of both species exhibited greater sensitivity to UV-B exposure. Older larvae of A. maculatum that were not exposed to UV-B as embryos were more sensitive than larvae that had hatched during exposure to UV-B. Growth of surviving larvae of A. maculatum was significantly reduced as UV-B intensity increased, whereas growth of A. talpoideum was unaffected. These results were compared to ambient UV-B conditions in natural environments. It appears that the embryo stage is relatively unaffected by UV-B levels observed in natural habitats, probably because of protection from vegetation, organic matter in the water column, oviposition depth, and egg jelly. The larval stage of these species may be at greater risk, particularly if there is an increase in UV-B radiation exposure caused by increases in water clarity and/or decreases in dissolved organic carbon.
Garcia, T.S.; Straus, R.; Sih, A.
Temperature has been shown to affect body color in several species of amphibians. The interaction between color and temperature may also change over larval ontogeny, perhaps because of age-related or seasonal changes in selection pressures on color. We quantified the effects of temperature on the color of the salamander sister species Ambystoma barbouri and Ambystoma texanum over larval ontogeny. We found that early-stage larvae responded to cold temperatures with a dark color relative to that of the warm temperature response. Both species then exhibited an ontogenetic shift in larval color, with larvae becoming lighter with age. Interestingly, older larvae showed decreased plasticity in color change to temperature when compared with younger stages. Older A. texanum larvae exhibited a reversal in the direction of color change, with cold temperatures inducing a lighter color relative to warm temperatures. We suggest that the overall pattern of color change (a plastic color response to temperature for young larvae, a progressive lightening of larvae over development, and an apparent loss of color plasticity to temperature over ontogeny) can be plausibly explained by seasonal changes in environmental factors (temperature, ultraviolet radiation) selecting for body color. (author)
Glorioso, Brad M.; Waddle, J. Hardin; Hefner, J. M.
Reproductive aspects of Ambystoma salamanders were investigated at sites in Louisiana (2010–12) and Mississippi (2013). Three species occurred at the Louisiana site, Spotted Salamander (A. maculatum), Marbled Salamander (A. opacum), and Mole Salamander (A. talpoideum), whereas only Spotted Salamanders were studied at the Mississippi site. A total of 162 and 71 egg masses of Spotted Salamanders were examined at the Louisiana and Mississippi sites, respectively. Significantly more Spotted Salamander eggs per egg mass were observed at the Mississippi site (x̄ = 78.2) than the Louisiana site (x̄ = 53.8; P < 0.001). The mean snout–vent length of female Spotted Salamanders at the Mississippi site (82.9 mm) was significantly larger than the Louisiana site (76.1 mm; P < 0.001). Opaque Spotted Salamander egg masses were not found at the Mississippi site, but accounted for 11% of examined egg masses at the Louisiana site. The mean number of eggs per egg mass at the Louisiana site did not differ between opaque (47.3) and clear (54.6) egg masses (P = 0.21). A total of 47 egg masses of the Mole Salamander were examined, with a mean number of 6.7 embryos per mass. Twenty-three individual nests of the Marbled Salamander were found either under or in decaying logs in the dry pond basins. There was no difference between the mean numbers of eggs per mass of attended nests (93.0) versus those that were discovered unattended (86.6; P = 0.67). Females tended to place their nests at intermediate heights within the pond basin.
McMenamin Sarah K
Full Text Available Abstract Background Loss of pond habitat is catastrophic to aquatic larval amphibians, but even reduction in the amount of time a breeding site holds water (hydroperiod can influence amphibian development and limit reproductive success. Using the landscape variation of a glacial valley in the Greater Yellowstone Ecosystem as the context for a natural experiment, we examined variation in growth pattern and life history of the salamander Ambystoma tigrinum melanostictum and determined how these developmental characteristics varied with hydroperiod over several summers. Results In ponds that dried early in the season, maximum larval size was reduced relative to the sizes achieved in permanent ponds. Ephemeral ponds were associated with early metamorphosis at small body sizes, while permanent ponds facilitated longer larval periods and later metamorphosis. Paedomorphosis resulted from indefinite metamorphic postponement, and was identified only in the most permanent environments. Patterns of growth and allometry were similar between ponds with different hydroperiods, but considerable life history variation was derived from modulating the timing of and size at metamorphosis. Considering maximum rates of growth and inferring the minimum size at metamorphosis across 25 ponds over the course of three years, we calculated that hydroperiods longer than three months are necessary to support these populations through metamorphosis and/or reproductive maturity. Conclusions Landscape heterogeneity fosters life history variation in this natural population. Modulation of the complex ambystomatid life cycle allows this species to survive in unpredictable environments, but current trends towards rapid pond drying will promote metamorphosis at smaller sizes and could eliminate the paedomorphic phenotype from this region. Metamorphosis at small size is has been linked to altered fitness traits, including reduced survival and fecundity. Thus, widespread
Full Text Available Abstract Background Amphibians are sensitive indicators of environmental conditions and show measurable responses, such as changes in phenology, abundance and range limits to local changes in precipitation and temperature regimes. Amphibians offer unique opportunities to study the important ecological and evolutionary implications of responses in life history characteristics to climatic change. We analyzed a late-Holocene fossil record of the Tiger Salamander (Ambystoma tigrinum for evidence of population-level changes in body size and paedomorphosis to climatic change over the last 3000 years. Results We found a significant difference in body size index between paedomorphic and metamorphic individuals during the time interval dominated by the Medieval Warm Period. There is a consistent ratio of paedomorphic to metamorphic specimens through the entire 3000 years, demonstrating that not all life history characteristics of the population were significantly altered by changes in climate on this timescale. Conclusion The fossil record of Ambystoma tigrinum we used spans an ecologically relevant timescale appropriate for understanding population and community response to projected climatic change. The population-level responses we documented are concordant with expectations based on modern environmental studies, and yield insight into population-level patterns across hundreds of generations, especially the independence of different life history characteristics. These conclusions lead us to offer general predictions about the future response of this species based on likely scenarios of climatic warming in the Rocky Mountain region.
Bruzgul, Judsen E; Long, Webb; Hadly, Elizabeth A
Amphibians are sensitive indicators of environmental conditions and show measurable responses, such as changes in phenology, abundance and range limits to local changes in precipitation and temperature regimes. Amphibians offer unique opportunities to study the important ecological and evolutionary implications of responses in life history characteristics to climatic change. We analyzed a late-Holocene fossil record of the Tiger Salamander (Ambystoma tigrinum) for evidence of population-level changes in body size and paedomorphosis to climatic change over the last 3000 years. We found a significant difference in body size index between paedomorphic and metamorphic individuals during the time interval dominated by the Medieval Warm Period. There is a consistent ratio of paedomorphic to metamorphic specimens through the entire 3000 years, demonstrating that not all life history characteristics of the population were significantly altered by changes in climate on this timescale. The fossil record of Ambystoma tigrinum we used spans an ecologically relevant timescale appropriate for understanding population and community response to projected climatic change. The population-level responses we documented are concordant with expectations based on modern environmental studies, and yield insight into population-level patterns across hundreds of generations, especially the independence of different life history characteristics. These conclusions lead us to offer general predictions about the future response of this species based on likely scenarios of climatic warming in the Rocky Mountain region.
Hansen, G N; Hansen, B L; Jørgensen, P N
The pancreas of the axolotl, Ambystoma mexicanum, was investigated by immunocytochemical methods for the presence of immunoreactivity to a number of antisera raised against mammalian insulins. All anti-insulin antisera tested revealed substantial amounts of reaction products confined solely to th...
Hossack, Blake R.; Muths, Erin L.; Rorabaugh, James C.; Lemos Espinal, Julio A.; Sigafus, Brent H.; Chambert, Thierry A.; Carreon Arroyo, Gerardo; Hurtado Felix, David; Toyos Martinez, Daniel; Jones, Thomas R.
The Sonoran Tiger Salamander (Ambystoma mavortium stebbinsi Lowe, 1954) was listed as federally endangered in the USA in 1997 (USFWS 1997). In the USA, the distribution of A. mavortium stebbinsi is limited to the San Rafael Valley (approximately 567 km2), between the Sierra San Antonio (called the Patagonia Mountains in Arizona) and Huachuca Mountains, and south of the Canelo Hills, Arizona (Fig. 1). The USA listing was triggered by loss of natural wetland habitats, threats from invasive predators, frequent die-offs from disease, introgression with the introduced Barred Tiger Salamander (A. mavortium mavortium), and small range and number of breeding sites that increases susceptibility to stochastic events (USFWS 1997). Small population sizes and limited gene flow have caused inbreeding, which may further reduce population viability and the potential for recovery (Jones et al. 1988; Storfer et al. 2014).
Takami, Yoshinori; Une, Yumi
Ambystoma mexicanum kept as pets are affected by a variety of diseases. However, no reports regarding the incidence of specific diseases are available. This study aimed to identify the diseases that occur frequently in this species by surveying the incidence of conditions in pet A. mexicanum specimens brought to a veterinary hospital. The sample comprised 97 pet A. mexicanum individuals brought to the authors' hospital during the 82-month period, i.e., from January 2008 to October 2014. In total, 116 diseases were identified. The most common disease was hydrocoelom (32 cases; 27.5% of all cases). Elucidating the pathogenesis of hydrocoelom, which has a high prevalence rate, is vital to maintaining the long-term health of A. mexicanum pets.
Hoffman, Robert L.; Larson, Gary L.; Samora, B.
Introduced, nonnative brook trout (Salvelinus fontinalis) were removed from a mountain lake in Mount Rainier National Park, Washington, to examine the capacity of native Ambystoma gracile (Northwestern Salamander) in the lake to respond to the intentional removal of fish. Temporal trends (N) were calculated for A. gracile larvae/neotene and egg mass relative abundances in the Fish Removal and an adjacent Fishless Lake. The diel and spatial patterns of A. gracile in the lakes were also enumerated during time-intervals of fish presence in and after fish removal from the Fish Removal Lake. Sixty-six fish were removed from the Fish Removal Lake. The Ns for relative abundances in the Fish Removal Lake were positive for the study period and indicated that the number of larvae/neotenes and egg masses observed in the lake increased concurrent with the removal and extirpation of fish from the lake. Numbers of larvae/neotenes and egg masses observed in the Fishless Lake varied annually, but no overall positive or negative trends were evident during the study. Ambystoma gracile in the Fish Removal Lake, during fish presence, were predominantly nocturnal and located in the shallow, structurally complex nearshore area of the lake. After fish were removed, the number of A. gracile observed in the lake increased, especially during the day and in the deeper, less structurally complex offshore area of the lake. Fishless Lake A. gracile were readily observed day and night in all areas of the lake throughout the study. The A. gracile in the Fish Removal Lake behaviorally adapted to the presence of introduced fish and were able to recover from the affects of the fish following fish removal. This study underscores the important relationship between species life history and the variability of responses of montane aquatic-breeding amphibians to fish introductions in mountain lakes.
Diego de Jesus Chaparro-Herrera
Full Text Available Ambystoma mexicanum, a highly endangered species, is endemic to lake Xochimilco (Mexico City, Mexico which currently is being negatively affected by the introduction of Oreochromis niloticus (Tilapia and water pollution. During the first weeks of development, when mortality is the highest, Ambystoma mexicanumdepends on a diet of zooplankton. The aim of this study was to check whether contamination levels in lake Xochimilco influence zooplankton consumption by similar size classes of A. mexicanum and Oreochromis niloticus. In this study, we analysed changes in the functional responses and prey preference of A. mexicanum and larval Tilapia in two media, one with filtered lake Xochimilco water and another one with reconstituted water. As prey we used cladocerans (Moina macrocopa, Alona glabra, Macrothrix triserialis and Simocephalus vetulus and ostracods (Heterocypris incongruens. Zooplankton was offered in 5 different densities, 10, 20, 40, 80, 160 ind./mL. Prey consumption by A. mexicanum varied in relation to the species offered and age of the larvae. From the first week to the eighth week prey consumption by A. mexicanum increased by 57%. Our functional response tests showed that regardless of the prey type, prey consumption by A. mexicanum was lower in the contaminated water from lake Xochimilco. Among the zooplankton offered in the contaminated environment predators preferred smaller and slower moving microcrustaceans such as Alona glabra and Heterocypris incongruens. Furthermore, O. niloticus preferred prey such as Moina macrocopa and Macrothrix triserialis in the contaminated medium and was more voracious than the axolotl. Our results indicate that both water quality of the lake and the presence of the more resistant exotic fish adversely impact the survival of this endangered amphibian.
Khattak, Shahryar; Murawala, Prayag; Andreas, Heino; Kappert, Verena; Schuez, Maritta; Sandoval-Guzmán, Tatiana; Crawford, Karen; Tanaka, Elly M
The axolotl (Mexican salamander, Ambystoma mexicanum) has become a very useful model organism for studying limb and spinal cord regeneration because of its high regenerative capacity. Here we present a protocol for successfully mating and breeding axolotls in the laboratory throughout the year, for metamorphosing axolotls by a single i.p. injection and for axolotl transgenesis using I-SceI meganuclease and the mini Tol2 transposon system. Tol2-mediated transgenesis provides different features and advantages compared with I-SceI-mediated transgenesis, and it can result in more than 30% of animals expressing the transgene throughout their bodies so that they can be directly used for experimentation. By using Tol2-mediated transgenesis, experiments can be performed within weeks (e.g., 5-6 weeks for obtaining 2-3-cm-long larvae) without the need to establish germline transgenic lines (which take 12-18 months). In addition, we describe here tamoxifen-induced Cre-mediated recombination in transgenic axolotls.
Full Text Available BACKGROUND: A major step during the evolution of tetrapods was their transition from water to land. This process involved the reduction or complete loss of the dermal bones that made up connections to the skull and a concomitant enlargement of the endochondral shoulder girdle. In the mouse the latter is derived from three separate embryonic sources: lateral plate mesoderm, somites, and neural crest. The neural crest was suggested to sustain the muscle attachments. How this complex composition of the endochondral shoulder girdle arose during evolution and whether it is shared by all tetrapods is unknown. Salamanders that lack dermal bone within their shoulder girdle were of special interest for a possible contribution of the neural crest to the endochondral elements and muscle attachment sites, and we therefore studied them in this context. RESULTS: We grafted neural crest from GFP+ fluorescent transgenic axolotl (Ambystoma mexicanum donor embryos into white (d/d axolotl hosts and followed the presence of neural crest cells within the cartilage of the shoulder girdle and the connective tissue of muscle attachment sites of the neck-shoulder region. Strikingly, neural crest cells did not contribute to any part of the endochondral shoulder girdle or to the connective tissue at muscle attachment sites in axolotl. CONCLUSIONS: Our results in axolotl suggest that neural crest does not serve a general function in vertebrate shoulder muscle attachment sites as predicted by the "muscle scaffold theory," and that it is not necessary to maintain connectivity of the endochondral shoulder girdle to the skull. Our data support the possibility that the contribution of the neural crest to the endochondral shoulder girdle, which is observed in the mouse, arose de novo in mammals as a developmental basis for their skeletal synapomorphies. This further supports the hypothesis of an increased neural crest diversification during vertebrate evolution.
Busse, U; Séguin, C
To analyze Wnt-1 expression during neurulation in urodele embryos, we have isolated a Wnt-1 cDNA clone, Awnt-1, from an Ambystoma mexicanum (axolotl) neurula-stage cDNA library. Awnt-1 codes for a protein of 369 amino acids rich in cysteine residues, is preceded by a hydrophobic leader peptide sequence and contains four possible sites for N-linked glycosylation. The temporal expression profile of Awnt-1 was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Awnt-1 expression in the axolotl embryo is biphasic. Awnt-1 transcripts are found in early blastulae until gastrulation, are barely detectable during gastrulation, and are present again from neurulation until late embryogenesis. Transcripts are present before the midblastula transition, indicating that they might be of maternal origin. To localize Awnt-1 expression in embryos during the first phase of expression, early gastrulae were dissected by cutting along the animal-vegetal and future dorso-ventral axes and analyzed by RT-PCR. At the early gastrula stage Awnt-1 transcripts appear to be located in the future ventral region of the embryo. Hatching larvae no longer express Awnt-1. PCR reactions performed using cDNA library-phage DNA templates derived from whole neurulae versus embryos with the neuroectoderm removed suggest that, in the neurula, Awnt-1 transcripts are located in the neuroectoderm. This suggest that, as is the case for Wnt-1 in other vertebrates, Awnt-1 may be involved in neurogenesis. These results suggest that Wnt-1 has earlier roles in development than has been considered until now.
Jancovich, James K; Jacobs, Bertram L
Ranaviruses (family Iridoviridae, genus Ranavirus) are large, double-stranded DNA (dsDNA) viruses whose replication is restricted to ectothermic vertebrates. Many highly pathogenic members of the genus Ranavirus encode a homologue of the eukaryotic translation initiation factor 2α (eIF2α). Data in a heterologous vaccinia virus system suggest that the Ambystoma tigrinum virus (ATV) eIF2α homologue (vIF2αH; open reading frame [ORF] 57R) is involved in evading the host innate immune response by degrading the interferon-inducible, dsRNA-activated protein kinase, PKR. To test this hypothesis directly, the ATV vIF2αH gene (ORF 57R) was deleted by homologous recombination, and a selectable marker was inserted in its place. The ATVΔ57R virus has a small plaque phenotype and is 8-fold more sensitive to interferon than wild-type ATV (wtATV). Infection of fish cells with the ATVΔ57R virus leads to eIF2α phosphorylation, in contrast to infection with wtATV, which actively inhibits eIF2α phosphorylation. The inability of ATVΔ57R to prevent phosphorylation of eIF2α correlates with degradation of fish PKZ, an interferon-inducible enzyme that is closely related to mammalian PKR. In addition, salamanders infected with ATVΔ57R displayed an increased time to death compared to that of wtATV-infected salamanders. Therefore, in a biologically relevant system, the ATV vIF2αH gene acts as an innate immune evasion factor, thereby enhancing virus pathogenesis.
Julio A. Lemos-Espinal
Full Text Available Several species of salamander in the genus Ambystoma occur in the mountains surrounding Mexico City and are considered at risk of extinction. However, little is known about their ecology and natural history. The Toluca Stream Siredon (Ambystoma rivulare is classified as “Data Deficient” by the IUCN, and considered “Threatened” under Mexican law. From October 2013 to September 2014, we examined the diet of larval A. rivulare from a stream on the Volcán Nevado de Toluca in Mexico to provide insight into the suitability of the habitat to support this population of salamanders. Ostracods accounted for approximately 90% of all prey items consumed by larval A. rivulare. The number of ostracods found in stomachs increased with individual body size, but the proportion of ostracods in stomachs did not vary with body size. Nematodes were observed in approximately one third of the stomachs we examined. The diversity of prey in the diet of A. rivulare in the stream we studied is low and dominated by a single prey taxon, ostracods. Our results suggest that if environmental conditions in the stream change such that ostracods are negatively affected then the long-term persistence of this population of A. rivulare might be in jeopardy.
Currens, C.R.; Liss, W.J.; Hoffman, R.L.
The formation of amphibian population structure is directly affected by predation. Although aquatic predators have been shown to have direct negative effects on larval salamanders in laboratory and field experiments, the potential impacts of gape-limited fish on larval salamander growth has been largely underexplored. We designed an enclosure experiment conducted in situ to quantify the effects of gape-limited Brook Trout (Salvelinus fontinalis) on larval Northwestern Salamander (Ambystoma gracile) growth. We specifically tested whether the presence of fish too small to consume larvae had a negative effect on larval growth. The results of this study indicate that the presence of a gape-limited S. fontinalis can have a negative effect on growth of larval A. gracile salamanders. Copyright 2007 Society for the Study of Amphibians and Reptiles.
Baxter, Leilan; Brain, Richard A.; Hosmer, Alan J.; Nema, Mohini; Müller, Kirsten M.; Solomon, Keith R.; Hanson, Mark L.
Embryonic growth of the yellow-spotted salamander (Ambystoma maculatum) is enhanced by the presence of the green alga Oophila amblystomatis, in the egg capsule. To further assess potential impacts of herbicides on this relationship, A. maculatum egg masses were exposed to atrazine (0–338 μg/L) until hatching (up to 66 days). Exposure to atrazine reduced PSII yield of the symbiotic algae in a concentration-dependent manner, but did not significantly affect visible algal growth or any metrics associated with salamander development. Algal cells were also cultured in the laboratory for toxicity testing. In the 96-h growth inhibition test (0–680 μg/L), ECx values were generally greater than those reported for standard algal test species. Complete recovery of growth rates occurred within 96-h of transferring cells to untreated media. Overall, development of A. maculatum embryos was not affected by exposure to atrazine at concentrations and durations exceeding those found in the environment. - Highlights: • The yellow-spotted salamander produces eggs that are colonized by a symbiotic green alga. • We tested the sensitivity of this system to the herbicide atrazine. • Embryo development was not significantly affected by exposure at up to 300 μg/L. • The alga was isolated and 96-h growth tests were performed in the laboratory. • EC50s for Oophila sp. were >100 μg/L. - Development of Ambystoma maculatum embryos in egg masses was not impacted by exposure to atrazine at concentrations and durations exceeding those commonly found in the environment.
Calfee, Robin D.; Little, Edward E.; Pearl, Christopher A.; Hoffman, Robert L.
Solar ultraviolet radiation (UV) has received much attention as a factor that could play a role in amphibian population declines. UV can be hazardous to some amphibians, but the resultant effects depend on a variety of environmental and behavioral factors. In this study, the potential effects of UV on the Northwestern Salamander, Ambystoma gracile, from three lakes were assessed in the laboratory using a solar simulator. We measured the survival of embryos and the survival and growth of larvae exposed to four UV treatments in controlled laboratory studies, the UV absorbance of egg jelly, oviposition depths in the lakes, and UV absorbance in water samples from the three lakes. Hatching success of embryos decreased in the higher UV treatments as compared to the control treatments, and growth of surviving larvae was significantly reduced in the higher UVB irradiance treatments. The egg jelly exhibited a small peak of absorbance within the UVB range (290–320 nm). The magnitude of UV absorbance differed among egg jellies from the three lakes. Oviposition depths at the three sites averaged 1.10 m below the water surface. Approximately 66% of surface UVB radiation was attenuated at 10-cm depth in all three lakes. Results of this study indicate that larvae may be sensitive to UVB exposure under laboratory conditions; however, in field conditions the depths of egg deposition in the lakes, absorbance of UV radiation by the water column, and the potential for behavioral adjustments may mitigate severe effects of UV radiation.
Yu, Haining; Gao, Jiuxiang; Lu, Yiling; Guang, Huijuan; Cai, Shasha; Zhang, Songyan; Wang, Yipeng
Lysozymes are key proteins that play important roles in innate immune defense in many animal phyla by breaking down the bacterial cell-walls. In this study, we report the molecular cloning, sequence analysis and phylogeny of the first caudate amphibian g-lysozyme: a full-length spleen cDNA library from axolotl (Ambystoma mexicanum). A goose-type (g-lysozyme) EST was identified and the full-length cDNA was obtained using RACE-PCR. The axolotl g-lysozyme sequence represents an open reading frame for a putative signal peptide and the mature protein composed of 184 amino acids. The calculated molecular mass and the theoretical isoelectric point (pl) of this mature protein are 21523.0 Da and 4.37, respectively. Expression of g-lysozyme mRNA is predominantly found in skin, with lower levels in spleen, liver, muscle, and lung. Phylogenetic analysis revealed that caudate amphibian g-lysozyme had distinct evolution pattern for being juxtaposed with not only anura amphibian, but also with the fish, bird and mammal. Although the first complete cDNA sequence for caudate amphibian g-lysozyme is reported in the present study, clones encoding axolotl's other functional immune molecules in the full-length cDNA library will have to be further sequenced to gain insight into the fundamental aspects of antibacterial mechanisms in caudate.
Calfee, R.D.; Little, E.E.; Pearl, C.A.; Hoffman, R.L.
Solar ultraviolet radiation (UV) has received much attention as a factor that could play a role in amphibian population declines. UV can be hazardous to some amphibians, but the resultant effects depend on a variety of environmental and behavioral factors. In this study, the potential effects of UV on the Northwestern Salamander, Ambystoma gracile, from three lakes were assessed in the laboratory using a solar simulator. We measured the survival of embryos and the survival and growth of larvae exposed to four UV treatments in controlled laboratory studies, the UV absorbance of egg jelly, oviposition depths in the lakes, and UV absorbance in water samples from the three lakes. Hatching success of embryos decreased in the higher UV treatments as compared to the control treatments, and growth of surviving larvae was significantly reduced in the higher UVB irradiance treatments. The egg jelly exhibited a small peak of absorbance within the UVB range (290-320 nm). The magnitude of UV absorbance differed among egg jellies from the three lakes. Oviposition depths at the three sites averaged 1.10 m below the water surface. Approximately 66 of surface UVB radiation was attenuated at 10-cm depth in all three lakes. Results of this study indicate that larvae may be sensitive to UVB exposure under laboratory conditions; however, in field conditions the depths of egg deposition in the lakes, absorbance of UV radiation by the water column, and the potential for behavioral adjustments may mitigate severe effects of UV radiation. Copyright 2010 Society for the Study of Amphibians and Reptiles.
Norman, M.F.; Norris, D.O.
The sensitivity of thyroid glands from the tiger salamander, Ambystoma tigrinum, to bovine thyrotropin (bTSH) was tested in vitro. Thyroids were taken from subjects representing metamorphic stages I (premetamorphic larvae), II (onset of climax), and VII (completion of gill resorption), as well as from captivity control larvae. Exogenous TSH reduced the cumulative uptake of 125 I in vitro by thyroids from stage I larvae after 24 and 48 hr. The capacity of thyroids to release thyroxine (T4) in vitro was used subsequently as a measure of their responsiveness to TSH. Baseline levels of T4 release in vitro were variable but did not differ significantly among developmental stages. A low dose of bTSH (5 X 10(-6) IU/ml) did not increase in vitro T4 release compared with that of controls. A larger dose (5 X 10(-4) IU/ml) caused greater increases in T4 release from thyroids of stage II and VII subjects than from those of controls. This dose produced only a small response by thyroids from captivity-control subjects. The results suggest that the thyroids of Ambystoma increase in their capacity to respond to TSH during the process of metamorphosis
Kenison, Erin K; Litt, Andrea R.; Pilliod, David S.; McMahon, Tom E
Predation by nonnative fishes has reduced abundance and increased extinction risk for amphibian populations worldwide. Although rare, fish and palatable amphibians have been observed to coexist where aquatic vegetation and structural complexity provide suitable refugia. We examined whether larval long-toed salamanders (Ambystoma macrodactylum Baird, 1849) increased use of vegetation cover in lakes with trout and whether adding vegetation structure could reduce predation risk and nonconsumptive effects (NCEs), such as reductions in body size and delayed metamorphosis. We compared use of vegetation cover by larval salamanders in lakes with and without trout and conducted a field experiment to investigate the influence of added vegetation structure on salamander body morphology and life history. The probability of catching salamanders in traps in lakes with trout was positively correlated with the proportion of submerged vegetation and surface cover. Growth rates of salamanders in enclosures with trout cues decreased as much as 85% and the probability of metamorphosis decreased by 56%. We did not find evidence that adding vegetation reduced NCEs in experimental enclosures, but salamanders in lakes with trout utilized more highly-vegetated areas which suggests that adding vegetation structure at the scale of the whole lake may facilitate coexistence between salamanders and introduced trout.
Micheletti, Steven J; Storfer, Andrew
The central-marginal hypothesis (CMH) predicts that population size, genetic diversity and genetic connectivity are highest at the core and decrease near the edges of species' geographic distributions. We provide a test of the CMH using three replicated core-to-edge transects that encompass nearly the entire geographic range of the endemic streamside salamander (Ambystoma barbouri). We confirmed that the mapped core of the distribution was the most suitable habitat using ecological niche modelling (ENM) and via genetic estimates of effective population sizes. As predicted by the CMH, we found statistical support for decreased genetic diversity, effective population size and genetic connectivity from core to edge in western and northern transects, yet not along a southern transect. Based on our niche model, habitat suitability is lower towards the southern range edge, presumably leading to conflicting core-to-edge genetic patterns. These results suggest that multiple processes may influence a species' distribution based on the heterogeneity of habitat across a species' range and that replicated sampling may be needed to accurately test the CMH. Our work also emphasizes the importance of identifying the geographic range core with methods other than using the Euclidean centre on a map, which may help to explain discrepancies among other empirical tests of the CMH. Assessing core-to-edge population genetic patterns across an entire species' range accompanied with ENM can inform our general understanding of the mechanisms leading to species' geographic range limits. © 2015 John Wiley & Sons Ltd.
Full Text Available In spite of numerous investigations of regenerating salamander limbs, little attention has been paid to the details of how joints are reformed. An understanding of the process and mechanisms of joint regeneration in this model system for tetrapod limb regeneration would provide insights into developing novel therapies for inducing joint regeneration in humans. To this end, we have used the axolotl (Mexican Salamander model of limb regeneration to describe the morphology and the expression patterns of marker genes during joint regeneration in response to limb amputation. These data are consistent with the hypothesis that the mechanisms of joint formation whether it be development or regeneration are conserved. We also have determined that defects in the epiphyseal region of both forelimbs and hind limbs in the axolotl are regenerated only when the defect is small. As is the case with defects in the diaphysis, there is a critical size above which the endogenous regenerative response is not sufficient to regenerate the joint. This non-regenerative response in an animal that has the ability to regenerate perfectly provides the opportunity to screen for the signaling pathways to induce regeneration of articular cartilage and joints.
Phan, Anne Quy
Salamanders are unique among adult vertebrates in their ability to regenerate complex body structures after traumatic injury. Axolotl limb regeneration is a stepwise sequence of three requisite processes: (1) scarless wound healing to generate a regenerative wound epithelium, (2) blastema formation by migration, proliferation and dedifferentiation to create a mass of multipotent regeneration-competent progenitor cells, and (3) induction of pattern formation by interaction of cells with opposi...
.... Fish and Wildlife Service. The species inhabits the lower Southeastern Coastal Plain from southern South Carolina to northern Florida, and westward through Georgia to extreme southwestern Alabama...
Secor, Stephen M; Boehm, Matthew
The past decade has witnessed a dramatic increase in studies of amphibian and reptile specific dynamic action (SDA). These studies have demonstrated that SDA, the summed energy expended on meal digestion and assimilation, is affected significantly by meal size, meal type, and body size and to some extent by body temperature. While much of this attention has been directed at anuran and reptile SDA, we investigated the effects of meal size, meal type, and body temperature on the postprandial metabolic responses and the SDA of the tiger salamander (Ambystoma tigrinum tigrinum). We also compared the SDA responses among six species of Ambystoma salamanders representing the breadth of Ambystoma phylogeny. Postprandial peaks in VO(2) and VO(2), duration of elevated metabolism, and SDA of tiger salamanders increased with the size of cricket meals (2.5%-12.5% of body mass). For A. tigrinum, as for other ectotherms, a doubling of meal size results in an approximate doubling of SDA, a function of equal increases in peak VO(2) and duration. For nine meal types of equivalent size (5% of body mass), the digestion of hard-bodied prey (crickets, superworms, mealworms, beetles) generated larger SDA responses than the digestion of soft-bodied prey (redworms, beetle larvae). Body temperature affected the profile of postprandial metabolism, increasing the peak and shortening the duration of the profile as body temperature increased. SDA was equivalent among three body temperatures (20 degrees, 25 degrees, and 30 degrees C) but decreased significantly at 15 degrees C. Comparatively, the postprandial metabolic responses and SDA of Ambystoma jeffersonianum, Ambystoma maculatum, Ambystoma opacum, Ambystoma talpoideum, Ambystoma texanum, and the conspecific Ambystoma tigrinum mavortium digesting cricket meals that were 5% of their body mass were similar (independent of body mass) to those of A. t. tigrinum. Among the six species, standard metabolic rate, peak postprandial VO(2), and SDA
We investigated the metapopulation structure of the California tiger salamander (Ambystoma californiense) using a combination of indirect and direct methods to evaluate two key requirements of modern metapopulation models: 1) that patches support somewhat independent populations ...
Vicksburg, MS. Pauly, G.B., Piskurek, O., Shaffer , H.B. 2007. Phylogeographic concordance in the southeastern United States: the flatwoods salamander...Flatwoods salamander (Ambystoma bishopi and A. cingulatum) Dataset: Pauly, G. B., Piskurek, O., Shaffer , H. B. 2007. Phylogeographic concordance in the...B., Piskurek, O., Shaffer , H. B. 2007. Phylogeographic concordance in the southeastern United States: the flatwoods salamander, Ambystoma cingulatum
an item) intelligence gathering. The new Tactical Post Blast Course would include an applied instruction facility, vehicular storage facility, and six...Reticulated flatwoods salamander Ambystoma bishopi E Reptiles Eastern indigo snake Drymarchon couperi T Green sea turtle Chelonia mydas E Kemp’s ridley
...] Proposed Safe Harbor Agreement for California Red-legged Frog, California Tiger Salamander, Smith's Blue...-legged frog (Rana draytonii) and California tiger salamander (Ambystoma californiense) and federally..., California tiger salamander, Smith's blue butterfly, and Yadon's piperia on the property subject to the...
The mole salamander, Ambystoma altamirani is a microendemic species, which is distributed incentral México, within the trans-Mexican volcanic belt, and is one of the most threatened species due to habitat destruction and the introduction of exotic species. Nine microsatellite markers were used to determine the genetic ...
inflow and surface water data of Big Stone Lake, October, 1967. *: 104. Todd, David Keith, The Water Encyclopedia , Water Information Center, Port...Mudpuppy Necturus maculosus Tiger Salamander Ambystoma tigrinu American Toad Bufo americanus Great Plains Toad Bufo cognatus Cricket Frog _’ Acris
Ardea herodias), crows (Corvus brachyrhynchos), ravens (Corvus corax), garter snakes (Thamnophis species), bullfrogs ( Rana catesbeiana ), California...Tiger Salamander Habitat Assessment at Travis Air Force Base, Solano County, California Rana Resources 2005 Results of First Year Special-Status...Athene cunicularia Burrowing owl CSC Known Rana aurora draytonii California red-legged frog FT Potential Ambystoma californiense California tiger
Ardea herodias), crows (Corvus brachyrhynchos), ravens (Corvus corax), garter snakes (Thamnophis species), bullfrogs ( Rana catesbeiana ), California...California Tiger Salamander Habitat Assessment at Travis Air Force Base, Solano County, California Rana Resources 2005 Results of First Year Special...clover FE Potential Animals Rana aurora draytonii California red-legged frog FT Potential Ambystoma californiense California tiger salamander FT
natural gas, sand and gravel, natural carbon dioxide, lime , and scoria (USAF, 2002b). 3.5 LAND USE Since its establishment in 1942, CAFB has...salamander (Ambystoma tigrinum), side- blotched lizard (Uta stansburiana), northern harrier (Circus cyaneus), ferruginous hawk, several species of
Home; Journals; Journal of Genetics. ARMANDO SUNNY. Articles written in Journal of Genetics. Volume 96 Issue 6 December 2017 pp 873-883 RESEARCH ARTICLE. Genetic variability and structure of an isolated population of Ambystoma altamirani , a mole salamander that lives in the mountains of one of the largest ...
Thomas, R. M.
Suggests and describes laboratory activities in which the Mexican axolotl (Ambystoma mexicanum Shaw) is used, including experiments in embryology and early development, growth and regeneration, neoteny and metamorphosis, genetics and coloration, anatomy and physiology, and behavior. Discusses care and maintenance of animals. (CS)
Thygesen, Mathias; Rasmussen, Mikkel Mylius; Madsen, Jesper Guldsmed
The Mexican axolotl (Ambystoma mexicanum) is an important model species in regenerative biology. Traditionally, axolotls are anesthetized using benzocaine or MS-222, both of which act to inhibit voltage gated sodium channels thereby preventing action potential propagation. In some neurophysiologi......The Mexican axolotl (Ambystoma mexicanum) is an important model species in regenerative biology. Traditionally, axolotls are anesthetized using benzocaine or MS-222, both of which act to inhibit voltage gated sodium channels thereby preventing action potential propagation. In some...... neurophysiological experiments this is not desirable; therefore we tested propofol as an alternative anesthetic in the axolotl. We evaluated benzocaine, MS-222, and propofol's cardiovascular effects, effects on action potential propagation in the spinal cord, and gross limb regenerative effects. We found...
von Byern, Janek; Mebs, Dietrich; Heiss, Egon; Dicke, Ursula; Wetjen, Oliver; Bakkegard, Kristin; Grunwald, Ingo; Wolbank, Susanne; Mühleder, Severin; Gugerell, Alfred; Fuchs, Heidemarie; Nürnberger, Sylvia
Salamanders have evolved a wide variety of antipredator mechanisms and behavior patterns, including toxins and noxious or adhesive skin secretions. The high bonding strength of the natural bioadhesives makes these substances interesting for biomimetic research and applications in industrial and medical sectors. Secretions of toxic species may help to understand the direct effect of harmful substances on the cellular level. In the present study, the biocompatibility of adhesive secretions from four salamander species (Plethodon shermani, Plethodon glutinosus, Ambystoma maculatum, Ambystoma opacum) were analyzed using the MTT assay in cell culture and evaluated against toxic secretions of Pleurodeles waltl, Triturus carnifex, Pseudotriton ruber, Tylototriton verrucosus, and Salamandra salamandra. Their effect on cells was tested in direct contact (direct culture) or under the influence of the extract (indirect exposure) in accordance with the protocol of the international standard norm ISO 10993-5. Human dermal fibroblasts (NHDF), umbilical vein endothelial cells (HUVEC), and articular chondrocytes (HAC), as well as the cell lines C2C12 and L929 were used in both culture types. While the adhesive secretions from Plethodon shermani are cytocompatible and those of Ambystoma opacum are even advantageous, those of Plethodon glutinosus and Ambystoma maculatum appear to be cytotoxic to NDHF and HUVEC. Toxic secretions from Salamandra salamandra exhibited harmful effects on all cell types. Pseudotriton ruber and Triturus carnifex secretions affected certain cell types marginally; those from Pleurodeles waltl and Tylototriton verrucosus were generally well tolerated. The study shows for the first time the effect of salamander secretions on the viability of different cell types in culture. Two adhesive secretions appeared to be cell compatible and are therefore promising candidates for future investigations in the field of medical bioadhesives. Among the toxic secretions
Zhu, Wei; Pao, Gerald M; Satoh, Akira; Cummings, Gillian; Monaghan, James R; Harkins, Timothy T; Bryant, Susan V; Voss, S Randal; Gardiner, David M; Hunter, Tony
The capacity for tissue and organ regeneration in humans is dwarfed by comparison to that of salamanders. Emerging evidence suggests that mechanisms learned from the early phase of salamander limb regeneration-wound healing, cellular dedifferentiation and blastemal formation-will reveal therapeutic approaches for tissue regeneration in humans. Here we describe a unique transcriptional fingerprint of regenerating limb tissue in the Mexican axolotl (Ambystoma mexicanum) that is indicative of ce...
Delgado Viveros, David
Esta región se ha considerado importante desde la época prehispánica, como un sistema alimentario y de amortiguamiento ambiental. Recursos naturales como Ambystoma mexicanum, la agricultura lacustre con Corisella texcocana y la agricultura de básicos, son aún fuente de alimento. Las políticas que se han aplicado influyeron en la desecación del lago. Debido a las consecuencias ecológicas de estas acciones se construyeron embalses artificiales, que mantuvieron los sistemas productivos aunque en...
Latney, La'Toya V; Klaphake, Eric
This review summarizes the most recent updates on emerging infectious diseases of amphibia. A brief summary of Batrachochytrium dendrobatidis history, epidemiology, pathogenesis, life cycle, diagnosis, treatment, and biosecurity is provided. Ambystoma tigrinum virus, common midwife toad virus, frog virus 3, Rana grylio virus, Rana catesbeiana ranavirus, Mahaffey Road virus, Rana esculenta virus, Bohle iridovirus, and tiger frog virus ranaviruses are extensively reviewed. Emerging bacterial pathogens are discussed, including Flavobacter sp, Aeromonas sp, Citrobacter freundii, Chlamydophila sp, Mycobacterium liflandii, Elizabethkingia meningoseptica, and Ochrobactrum anthropi. Rhabdias sp, Ribeiroia sp, and Spirometra erinacei are among several of the parasitic infections overviewed in this article. Copyright © 2013 Elsevier Inc. All rights reserved.
Richardson, Jonathan L; Urban, Mark C
Microgeographic adaptation occurs when populations evolve divergent fitness advantages across the spatial scales at which focal organisms regularly disperse. Although an increasing number of studies find evidence for microgeographic adaptation, the underlying causes often remain unknown. Adaptive divergence requires some combination of limited gene flow and strong divergent natural selection among populations. In this study, we estimated the relative influence of selection, gene flow, and the spatial arrangement of populations in shaping patterns of adaptive divergence in natural populations of the spotted salamander (Ambystoma maculatum). Within the study region, A. maculatum co-occur with the predatory marbled salamander (Ambystoma opacum) in some ponds, and past studies have established a link between predation risk and adaptive trait variation in A. maculatum. Using 14 microsatellite loci, we found a significant pattern of genetic divergence among A. maculatum populations corresponding to levels of A. opacum predation risk. Additionally, A. maculatum foraging rate was strongly associated with predation risk, genetic divergence, and the spatial relationship of ponds on the landscape. Our results indicate the sorting of adaptive genotypes by selection regime and strongly suggest that substantial selective barriers operate against gene flow. This outcome suggests that microgeographic adaptation in A. maculatum is possible because strong antagonistic selection quickly eliminates maladapted phenotypes despite ongoing and substantial immigration. Increasing evidence for microgeographic adaptation suggests a strong role for selective barriers in counteracting the homogenizing influence of gene flow. © 2013 The Author(s). Evolution © 2013 The Society for the Study of Evolution.
William B. Sutton
Full Text Available Global climate change represents one of the most extensive and pervasive threats to wildlife populations. Amphibians, specifically salamanders, are particularly susceptible to the effects of changing climates due to their restrictive physiological requirements and low vagility; however, little is known about which landscapes and species are vulnerable to climate change. Our study objectives included, (1 evaluating species-specific predictions (based on 2050 climate projections and vulnerabilities to climate change and (2 using collective species responses to identify areas of climate refugia for conservation priority salamanders in the northeastern United States. All evaluated salamander species were projected to lose a portion of their climatic niche. Averaged projected losses ranged from 3%–100% for individual species, with the Cow Knob Salamander (Plethodon punctatus, Cheat Mountain Salamander (Plethodon nettingi, Shenandoah Mountain Salamander (Plethodon virginia, Mabee’s Salamander (Ambystoma mabeei, and Streamside Salamander (Ambystoma barbouri predicted to lose at least 97% of their landscape-scale climatic niche. The Western Allegheny Plateau was predicted to lose the greatest salamander climate refugia richness (i.e., number of species with a climatically-suitable niche in a landscape patch, whereas the Central Appalachians provided refugia for the greatest number of species during current and projected climate scenarios. Our results can be used to identify species and landscapes that are likely to be further affected by climate change and potentially resilient habitats that will provide consistent climatic conditions in the face of environmental change.
Svistunov, S.A.; Mitashov, V.I.
Growth of the retina was studied in mature intact amphibians, tritons, axolotls, ambystomas and clawed frogs, for six months using multiple injection of 3 H-thymidine. It was established that the source of replenishment of the retina by new cells is its terminal zone in all animals investigated. This is attested to by the gradual migration of labeled cells from the growth zone into differentiated layers of the retina. The most intensely labeled cells occupy a distal position relative to other labeled cells, therefore marking the boundary between the initial part of the retina, not containing labeled nuclei, and the part being augmented. For each species studied, a level of proliferative activity is characteristic for cells of the terminal zone, which decreases in the order axolotl-clawed frog-triton -ambystoma. In the axolotl and additional growth zone is noted in the retina, in addition to the terminal, which is located in the area of the unclosed section of the embryonic fissure. Results obtained serve as a basis for the regenerative potentials of eye tissues revealed previously in these amphibian species
Semiltsch, Raymond D; Walls, Susan; Barichivich, William J.; O'Donnell, Katherine
A comprehensive view of population declines and their underlying causes is necessary to reverse species loss. Historically, in many cases, a narrow view may have allowed species declines to continue, virtually undetected, for long periods of time (perhaps even decades). We suggest that extinction debt is likely responsible for numerous (perhaps most) amphibian declines and that this perspective should be incorporated into the structure of amphibian research and management. Extinction debt, originally proposed to explain changes in species richness following environmental disturbance, also may refer to the proportion of populations of an individual species that is expected to eventually be lost because of habitat change. A conservation framework to address extinction debt focuses research on threats at the individual, population, and metapopulation levels. This approach will help enhance, restore, and protect specific processes and habitats at the proper scale by directing management to the most vulnerable level and stage of a species. We illustrate this approach using Flatwoods Salamanders, Ambystoma cingulatumand Ambystoma bishopi, which occurred historically throughout the Coastal Plain of the southeastern United States but have experienced a greater than 85% loss of populations in recent years. Reversal of these losses is possible only if conservation and recovery efforts encompass individual, population, and metapopulation levels. We illustrate our framework by outlining actions that could be taken at each of these levels to help guide conservation and management of amphibians with complex life cycles and provide options for how to prioritize conservation actions in the face of logistical and budgetary shortfalls.
Urban, Mark C
Ecological and evolutionary mechanisms are increasingly thought to shape local community dynamics. Here, I evaluate if the local adaptation of a meso-predator to an apex predator alters local food webs. The marbled salamander (Ambystoma opacum) is an apex predator that consumes both the spotted salamander (Ambystoma maculatum) and shared zooplankton prey. Common garden experiments reveal that spotted salamander populations which co-occur with marbled salamanders forage more intensely than those that face other predator species. These foraging differences, in turn, alter the diversity, abundance and composition of zooplankton communities in common garden experiments and natural ponds. Locally adapted spotted salamanders exacerbate prey biomass declines associated with apex predation, but dampen the top-down effects of apex predation on prey diversity. Countergradient selection on foraging explains why locally adapted spotted salamanders exacerbate prey biomass declines. The two salamander species prefer different prey species, which explains why adapted spotted salamanders buffer changes in prey composition owing to apex predation. Results suggest that local adaptation can strongly mediate effects from apex predation on local food webs. Community ecologists might often need to consider the evolutionary history of populations to understand local diversity patterns, food web dynamics, resource gradients and their responses to disturbance.
Zavala-Aguirre, Jose L; Torres-Bugarin, Olivia; Zamora-Perez, Ana L
A series of bioindicator organisms for aquatic ecosystems are being maintained under laboratory conditions in order to analyze effects of pollution on aquatic wildlife and potential effects on human health. Growth kinetics of algae Pseudokirchneriella subcapitata was used to evaluate effects of the surfactant nonilphenol (NP). Brachionus calyciflorus was used to set up a model of endocrine disruption using the fungicide vinclozolin (Vc). We exposed salamanders from the genus Ambystoma sp., to different concentrations of both the aneugen colchicine (COL) and the clastogen cyclophosphamide (CP) and we determined the frequency of micronucleated cells (MNC) in their shed skin. The presence of spontaneous micronuclei in peripheral blood erythrocytes from 10 fish species in Lake "La Alberca," Michoacan (Mexico), was evaluated as a possible biological indicator of genotoxic agents. Results confirm the sensivity of Pseudokirchneriella subcapitata to growth kinetics: the range of concentration of NP (20, 200 and 2000 microg L(- 1)) shows an inverted U shape in its maximum growth rate; Dimethyl sulfoxide (DMSO) used as a positive control and to solvate NP induced an inverse stimulatory effect on growth rate in the range of concentrations analyzed (0.0023, 0.023 and 0.23% v v(- 1)). In the use of the rotifer Brachionus calyciflorus, the range of Vc from 0.185 mg L(- 1) to 3 mg L(- 1) clearly showed an inverted U shape characteristic of endocrine disruptions. We were able to use shed skin from Ambystoma sp., to measure MNC frequencies induced either by an aneugenic or a clastogenic compound. The MNC frequency was increased significantly by all doses of COL and CP, administered either as single or repeated exposures. The presence of MNC in the shed skin and the speed of sloughing lead us to propose that the sheds of Ambystoma sp., or other amphibians that slough their skin, as suitable alternative models for detecting genotoxic exposures relevant to aquatic environments. In the
Chen, Guangchun; Robert, Jacques
Although a variety of virus species can infect amphibians, diseases caused by ranaviruses ([RVs]; Iridoviridae) have become prominent, and are a major concern for biodiversity, agriculture and international trade. The relatively recent and rapid increase in prevalence of RV infections, the wide range of host species infected by RVs, the variability in host resistance among population of the same species and among different developmental stages, all suggest an important involvement of the amphibian immune system. Nevertheless, the roles of the immune system in the etiology of viral diseases in amphibians are still poorly investigated. We review here the current knowledge of antiviral immunity in amphibians, focusing on model species such as the frog Xenopus and the salamander (Ambystoma tigrinum), and on recent progress in generating tools to better understand how host immune defenses control RV infections, pathogenicity, and transmission.
Lauridsen, Henrik; Foldager, Casper Bindzus; Hagensen, Mette
tissue without scar formation. Modern regenerative medicine seeks way to adopt these capacities to regenerative therapies in humans. Though much effort is put into the development of stem cell therapies, there exists currently no satisfying technique for non-invasive follow up examinations......Introduction: Regenerative potential in humans is very limited. Like other mammals we rely heavily on fibrosis and scar formation in response to injury. On the contrary, urodele amphibians (salamanders and newts) such as the axolotl (Ambystoma mexicanum) are champions of tissue regeneration among...... vertebrates mastering the ability to replace most tissues in addition to whole limbs, tail, jaw, etc. following damage or amputation. Regeneration in this species is taking place by dedifferentiation of cells to form a collection of stem cells, the regenerative blastema, that proliferate and regenerate lost...
Collins, Sara J; Russell, Ronald W
The deposition of chemical pollutants into roadside wetlands from runoff is a current environmental concern. In northern latitudes, a major pollutant in runoff water is salt (NaCl), used as de-icing agents. In this study, 26 roadside ponds were surveyed for amphibian species richness and chloride concentration. Acute toxicity tests (LC(50)) were performed on five locally common amphibian species using a range of environmentally significant NaCl concentrations. Field surveys indicated that spotted salamanders (Ambystoma maculatum) and wood frogs (Rana sylvatica) did not occupy high chloride ponds. American toads (Bufo americanus) showed no pond preference based on chloride concentration. Acute toxicity tests showed spotted salamanders and wood frogs were most sensitive to chloride, and American toads were the least. Spring peepers (Pseudacris crucifer) and green frogs (Rana clamitans) showed intermediate sensitivities. We concluded that chloride concentrations in ponds due to application of de-icing salts, influenced community structure by excluding salt intolerant species.
Hansen, Kasper; Lauridsen, Henrik; Pedersen, Michael
of stem cells), that proliferate and regenerate lost tissue completely without scar formation. The objective of this study was to evaluate if the positive effect of HBOT in wound healing translates to whole limb regeneration in a non-necrotic environment. The effect of two different constitutive HBOT......, whereas HBOT2 were pressurized in gaseous oxygen with 100% relative humidity. Control animals were exposed to normobaric conditions in water. Regeneration rate was evaluated as the increasing length of the regenerating appendage. Animals were kept at 20°C at all times. Results: Due to a group n = 2......: This pilot study revealed no effect of 80 days constitutive HBOT in a regeneration-competent species, suggesting that HBOT has minor consequences on tissue regeneration in a non-necrotic environment. Keywords: whole limb regeneration, amphibian, Mexican axolotl (Ambystoma mexicanum), HBOT...
McCay, T.S.; Forschler, B.T.; Komoroski, M.J.; Ford, W.M.
Sampled shrews at 9.3 ha plots from logs manually removed and control plots in loblolly pine forests of the Southeastern Coastal Plain. Capture rates of Cryptotis parva were lower at plots from which deadwood was removed whereas capture rates of Blarina cavolinensis and Sorex longirostris did not differ between control and removal plots. Cryptotis may have been most sensitive to removal plots due to low population density, hence poor ability to move into areas of low reproduction. (Second Abstract, p. 37)Presentation of evidence that juvenile amphibians including Ambystomatid salamanders may disperse hundreds of meter from their natal wetlands within the weeks to months following metamorphosis. Data indicates Ambystoma trigrinum metamorphs can take at least six months to disperse and en route use non-polar lipid reserves garnished as larvae. Report suggests a land management regime that allows for both juvenile amphibian dispersal and also the consumptive use of the surrounding landscape.
Haugan, Birgitte M; Halberg, Kenneth Agerlin; Jespersen, Åse
+ and K+ conductances, as well as a large K+ conductance in the basolateral cell membrane. Immunolabeling experiments indicate heavy expression of Na+/K+-ATPase in the basolateral labyrinth. Conclusions We propose that the pronephric duct is important for the subsequent modification of urine produced...... by the pronephros. Our results indicate that it reabsorbs sodium and secretes potassium via channels present in the apical cell membrane with the driving force for ion movement provided by the Na+/K+ pump. This is to our knowledge the first characterization of the pronephric duct, the precursor of the collecting...... whether the duct is involved in urine modification using larvae of the freshwater amphibian Ambystoma mexicanum (axolotl) as model. Results We investigated structural as well as physiological properties of the pronephric duct. The key elements of our methodology were: using histology, light...
Anderson, Thomas L; Rowland, Freya E; Semlitsch, Raymond D
Variation in the timing of breeding (i.e., phenological variation) can affect species interactions and community structure, in part by shifting body size differences between species. Body size differences can be further altered by density-dependent competition, though synergistic effects of density and phenology on species interactions are rarely evaluated. We tested how field-realistic variation in phenology and density affected ringed salamander (Ambystoma annulatum) predation on spotted salamanders (Ambystoma maculatum), and whether these altered salamander dynamics resulted in trophic cascades. In outdoor mesocosms, we experimentally manipulated ringed salamander density (low/high) and breeding phenology (early/late) of both species. Ringed salamander body size at metamorphosis, development, and growth were reduced at higher densities, while delayed phenology increased hatchling size and larval development, but reduced relative growth rates. Survival of ringed salamanders was affected by the interactive effects of phenology and density. In contrast, spotted salamander growth, size at metamorphosis, and survival, as well as the biomass of lower trophic levels, were negatively affected primarily by ringed salamander density. In an additional mesocosm experiment, we isolated whether ringed salamanders could deplete shared resources prior to their interactions with spotted salamanders, but instead found direct interactions (e.g., predation) were the more likely mechanism by which ringed salamanders limited spotted salamanders. Overall, our results indicate the effects of phenological variability on fitness-related traits can be modified or superseded by differences in density dependence. Identifying such context dependencies will lead to greater insight into when phenological variation will likely alter species interactions.
Bogart, J P; Bi, K
Polyploid animals have independently evolved from diploids in diverse taxa across the tree of life. We review a few polyploid animal species or biotypes where recently developed molecular and cytogenetic methods have significantly improved our understanding of their genetics, reproduction and evolution. Mitochondrial sequences that target the maternal ancestor of a polyploid show that polyploids may have single (e.g. unisexual salamanders in the genus Ambystoma) or multiple (e.g. parthenogenetic polyploid lizards in the genus Aspidoscelis) origins. Microsatellites are nuclear markers that can be used to analyze genetic recombinations, reproductive modes (e.g. Ambystoma) and recombination events (e.g. polyploid frogs such as Pelophylax esculentus). Hom(e)ologous chromosomes and rare intergenomic exchanges in allopolyploids have been distinguished by applying genome-specific fluorescent probes to chromosome spreads. Polyploids arise, and are maintained, through perturbations of the 'normal' meiotic program that would include pre-meiotic chromosome replication and genomic integrity of homologs. When possible, asexual, unisexual and bisexual polyploid species or biotypes interact with diploid relatives, and genes are passed from diploid to polyploid gene pools, which increase genetic diversity and ultimately evolutionary flexibility in the polyploid. When diploid relatives do not exist, polyploids can interact with another polyploid (e.g. species of African Clawed Frogs in the genus Xenopus). Some polyploid fish (e.g. salmonids) and frogs (Xenopus) represent independent lineages whose ancestors experienced whole genome duplication events. Some tetraploid frogs (P. esculentus) and fish (Squaliusalburnoides) may be in the process of becoming independent species, but diploid and triploid forms of these 'species' continue to genetically interact with the comparatively few tetraploid populations. Genetic and genomic interaction between polyploids and diploids is a complex
Full Text Available Abstract Background The Mexican axolotl (Ambystoma mexicanum is considered a hopeful monster because it exhibits an adaptive and derived mode of development - paedomorphosis - that has evolved rapidly and independently among tiger salamanders. Unlike related tiger salamanders that undergo metamorphosis, axolotls retain larval morphological traits into adulthood and thus present an adult body plan that differs dramatically from the ancestral (metamorphic form. The basis of paedomorphic development was investigated by comparing temporal patterns of gene transcription between axolotl and tiger salamander larvae (Ambystoma tigrinum tigrinum that typically undergo a metamorphosis. Results Transcript abundances from whole brain and pituitary were estimated via microarray analysis on four different days post hatching (42, 56, 70, 84 dph and regression modeling was used to independently identify genes that were differentially expressed as a function of time in both species. Collectively, more differentially expressed genes (DEGs were identified as unique to the axolotl (n = 76 and tiger salamander (n = 292 than were identified as shared (n = 108. All but two of the shared DEGs exhibited the same temporal pattern of expression and the unique genes tended to show greater changes later in the larval period when tiger salamander larvae were undergoing anatomical metamorphosis. A second, complementary analysis that directly compared the expression of 1320 genes between the species identified 409 genes that differed as a function of species or the interaction between time and species. Of these 409 DEGs, 84% exhibited higher abundances in tiger salamander larvae at all sampling times. Conclusions Many of the unique tiger salamander transcriptional responses are probably associated with metamorphic biological processes. However, the axolotl also showed unique patterns of transcription early in development. In particular, the axolotl showed a genome
Walls, Susan C.; Barichivich, William J.; Brown, Mary E.
The Class Amphibia is one of the most severely impacted taxa in an on-going global biodiversity crisis. Because amphibian reproduction is tightly associated with the presence of water, climatic changes that affect water availability pose a particularly menacing threat to both aquatic and terrestrial-breeding amphibians. We explore the impacts that one facet of climate change—that of extreme variation in precipitation—may have on amphibians. This variation is manifested principally as increases in the incidence and severity of both drought and major storm events. We stress the need to consider not only total precipitation amounts but also the pattern and timing of rainfall events. Such rainfall “pulses” are likely to become increasingly more influential on amphibians, especially in relation to seasonal reproduction. Changes in reproductive phenology can strongly influence the outcome of competitive and predatory interactions, thus potentially altering community dynamics in assemblages of co-existing species. We present a conceptual model to illustrate possible landscape and metapopulation consequences of alternative climate change scenarios for pond-breeding amphibians, using the Mole Salamander, Ambystoma talpoideum, as an example. Although amphibians have evolved a variety of life history strategies that enable them to cope with environmental uncertainty, it is unclear whether adaptations can keep pace with the escalating rate of climate change. Climate change, especially in combination with other stressors, is a daunting challenge for the persistence of amphibians and, thus, the conservation of global biodiversity. PMID:24832668
Galindo-Bustos, Miguel Angel; Hernandez-Jauregui, Dulce María Brousset; Cheng, Tina; Vredenburg, Vance; Parra-Olea, Gabriela
In Mexico City, native and exotic amphibians are commonly sold through the pet trade. This study investigates the presence of Batrachochytrium dendrobatidis (Bd) in native amphibians being sold at two commercial markets and at a herpetarium in Mexico City. A total of 238 individuals (6 genera and 12 species) were tested for Bd using real-time polymerase chain reaction (PCR) analysis. There were 197 Bd-positive individuals (prevalence 82%) from five species of amphibians. Hyla eximia from the markets had very high Bd prevalence (100%; 76/76 and 99%; 88/89) but those from the herpetarium were Bd negative (0/12). Ambystoma mexicanum from the herpetarium also had a high Bd-positive prevalence (80%; 28/35). Though A. mexicanum is nearly extinct in the wild, a commercial market continues to flourish through the pet trade. Now that captive colonies of A. mexicanum are currently used for reintroduction programs, the authors recommend quarantine to reduce spread of Bd via movement of infected animals in the trade and between colonies and via disposal of wastewater from captive collections.
Kluthe, Gregory J; Hillman, Stanley S
This study used an in situ heart preparation to analyze the power output and stroke work of spontaneously beating hearts of four anurans (Rhinella marina, Lithobates catesbeianus, Xenopus laevis, Pyxicephalus edulis) and three urodeles (Necturus maculosus, Ambystoma tigrinum, Amphiuma tridactylum) that span a representative range of relative ventricle mass (RVM) found in amphibians. Previous research has documented that RVM correlates with dehydration tolerance and maximal aerobic capacity in amphibians. The power output (mW g(-1) ventricle mass) and stroke work (mJ g(-1) ventricle muscle mass) were independent of RVM and were indistinguishable from previously published results for fish and reptiles. RVM was significantly correlated with maximum power output (P max, mW kg(-1) body mass), stroke volume, cardiac output, afterload pressure (P O) at P max, and preload pressure (P I) at P max. P I at P max and P O at P max also correlated very closely with each other. The increases in both P I and P O at maximal power outputs in large hearts suggest that concomitant increases in blood volume and/or increased modulation of vascular compliance either anatomically or via sympathetic tone on the venous vasculature would be necessary to achieve P max in vivo. Hypotheses for variation in RVM and its concomitant increased P max in amphibians are developed.
Voss, S R; Kump, D K; Walker, J A; Shaffer, H B; Voss, G J
The transformation of ancestral phenotypes into novel traits is poorly understood for many examples of evolutionary novelty. Ancestrally, salamanders have a biphasic life cycle with an aquatic larval stage, a brief and pronounced metamorphosis, followed by a terrestrial adult stage. Repeatedly during evolution, metamorphic timing has been delayed to exploit growth-permissive environments, resulting in paedomorphic salamanders that retain larval traits as adults. We used thyroid hormone (TH) to rescue metamorphic phenotypes in paedomorphic salamanders and then identified quantitative trait loci (QTL) for life history traits that are associated with amphibian life cycle evolution: metamorphic timing and adult body size. We demonstrate that paedomorphic tiger salamanders (Ambystoma tigrinum complex) carry alleles at three moderate effect QTL (met1-3) that vary in responsiveness to TH and additively affect metamorphic timing. Salamanders that delay metamorphosis attain significantly larger body sizes as adults and met2 explains a significant portion of this variation. Thus, substitution of alleles at TH-responsive loci suggests an adaptive pleiotropic basis for two key life-history traits in amphibians: body size and metamorphic timing. Our study demonstrates a likely pathway for the evolution of novel paedomorphic species from metamorphic ancestors via selection of TH-response alleles that delay metamorphic timing and increase adult body size.
Sanchez, Eugenia; Küpfer, Eliane; Goedbloed, Daniel J; Nolte, Arne W; Lüddecke, Tim; Schulz, Stefan; Vences, Miguel; Steinfartz, Sebastian
The postembryonic development of amphibians has been characterized as divided into three predominant periods, hereafter named primary developmental stages: premetamorphosis (PreM), prometamorphosis (ProM), metamorphic climax (Meta), and completion of metamorphosis (PostM), largely based on examination of anuran development. Here, we categorized the postembryonic development of larvae of a poisonous fire salamander (Salamandra salamandra) by integrating morphology and gene expression (transcriptomic) data. Morphological analysis revealed three distinct clusters suggestive of PreM, ProM, and Meta, which were confirmed in parallel by microarray-derived gene expression analysis. In total, 3,510 probes targeted transcripts differentially expressed between the clusters we identified. Genes upregulated in PreM related to organogenesis, and those upregulated in Meta underlie structural proteins and related to development of anatomical structures and pigmentation. Biosynthesis pathways of pigments (pteridines and melanin) were upregulated during late ProM and Meta. Gas chromatographic analysis of alkaloids indicated the onset of steroidal alkaloid biosynthesis at ProM. When comparing gene expression in the fire salamander to that in other amphibians-three anurans, Xenopus laevis, X. tropicalis, and Michrohyla fissipes, and one caudate, Ambystoma mexicanum- we identified genes with conserved expression patterns involved in basic metamorphic processes such as skin restructuring and tail fin resorption. Our results support that primary stages of postembryonic development in caudates are homologous to those of anurans, and offer a baseline for the study of the evolution of developmental modes. © 2018 Wiley Periodicals, Inc.
Stamm, Anne; Strauß, Sarah; Vogt, Peter; Scheper, Thomas; Pepelanova, Iliyana
AmbLOXe is a lipoxygenase, which is up-regulated during limb-redevelopment in the Mexican axolotl, Ambystoma mexicanum, an animal with remarkable regeneration capacity. Previous studies have shown that mammalian cells transformed with the gene of this epidermal lipoxygenase display faster migration and wound closure rate during in vitro wound healing experiments. In this study, the gene of AmbLOXe was codon-optimized for expression in Escherichia coli and was produced in the insoluble fraction as protein aggregates. These inclusion bodies or nanopills were shown to be reservoirs containing functional protein during in vitro wound healing assays. For this purpose, functional inclusion bodies were used to coat cell culture surfaces prior cell seeding or were added directly to the medium after cells reached confluence. In both scenarios, AmbLOXe inclusion bodies led to faster migration rate and wound closure, in comparison to controls containing either no AmbLOXe or GFP inclusion bodies. Our results demonstrate that AmbLOXe inclusion bodies are functional and may serve as stable reservoirs of this enzyme. Nevertheless, further studies with soluble enzyme are also necessary in order to start elucidating the exact molecular substrates of AmbLOXe and the biochemical pathways involved in the wound healing effect.
Zhu, Wei; Pao, Gerald M; Satoh, Akira; Cummings, Gillian; Monaghan, James R; Harkins, Timothy T; Bryant, Susan V; Voss, S Randal; Gardiner, David M; Hunter, Tony
The capacity for tissue and organ regeneration in humans is dwarfed by comparison to that of salamanders. Emerging evidence suggests that mechanisms learned from the early phase of salamander limb regeneration – wound healing, cellular dedifferentiation and blastemal formation – will reveal therapeutic approaches for tissue regeneration in humans. Here we describe a unique transcriptional fingerprint of regenerating limb tissue in the Mexican axolotl (Ambystoma mexicanum) that is indicative of cellular reprogramming of differentiated cells to a germline-like state. Two genes that are required for self-renewal of germ cells in mice and flies, Piwi-like 1 (PL1) and Piwi-like 2 (PL2), are expressed in limb blastemal cells, the basal layer keratinocytes and the thickened apical epithelial cap in the wound epidermis in the regenerating limb. Depletion of PL1 and PL2 by morpholino oligonucleotides decreased cell proliferation and increased cell death in the blastema leading to a significant retardation of regeneration. Examination of key molecules that are known to be required for limb development or regeneration further revealed that FGF8 is transcriptionally downregulated in the presence of the morpholino oligos, indicating PL1 and PL2 might participate in FGF signaling during limb regeneration. Given the requirement for FGF signaling in limb development and regeneration, the results suggest that PL1 and PL2 function to establish a unique germline-like state that is associated with successful regeneration. PMID:22841627
Demircan, Turan; İlhan, Ayşe Elif; Aytürk, Nilüfer; Yıldırım, Berna; Öztürk, Gürkan; Keskin, İlknur
Axolotl (Ambystoma Mexicanum) has been emerging as a promising model in stem cell and regeneration researches due to its exceptional regenerative capacity. Although it represents lifelong lasting neoteny, induction to metamorphosis with thyroid hormones (THs) treatment advances the utilization of Axolotl in various studies. It has been reported that amphibians undergo anatomical and histological remodeling during metamorphosis and this transformation is crucial for adaptation to terrestrial conditions. However, there is no comprehensive histological investigation regarding the morphological alterations of Axolotl organs and tissues throughout the metamorphosis. Here, we reveal the histological differences or resemblances between the neotenic and metamorphic axolotl tissues. In order to examine structural features and cellular organization of Axolotl organs, we performed Hematoxylin & Eosin, Luxol-Fast blue, Masson's trichrome, Alcian blue, Orcein and Weigart's staining. Stained samples from brain, gallbladder, heart, intestine, liver, lung, muscle, skin, spleen, stomach, tail, tongue and vessel were analyzed under the light microscope. Our findings contribute to the validation of the link between newly acquired functions and structural changes of tissues and organs as observed in tail, skin, gallbladder and spleen. We believe that this descriptive work provides new insights for a better histological understanding of both neotenic and metamorphic Axolotl tissues. Copyright © 2016 Elsevier GmbH. All rights reserved.
McLean, Kyle I.; Stockwell, Craig A.; Mushet, David M.
Barred tiger salamanders [Ambystoma mavortium (Baird, 1850)] exhibit two trophic morphologies; a typical and a cannibalistic morph. Cannibalistic morphs, distinguished by enlarged vomerine teeth, wide heads, slender bodies, and cannibalistic tendencies, are often found where conspecifics occur at high density. During 2012 and 2013, 162 North Dakota wetlands and lakes were sampled for salamanders. Fifty-one contained A. mavortium populations; four of these contained cannibalistic morph individuals. Two populations with cannibalistic morphs occurred at sites with high abundances of conspecifics. However, the other two populations occurred at sites with unexpectedly low conspecific but high fathead minnow [Pimephales promelas (Rafinesque, 1820)] abundances. Further, no typical morphs were observed in either of these later two populations, contrasting with earlier research suggesting cannibalistic morphs only occur at low frequencies in salamander populations. Another anomaly of all four populations was the occurrence of cannibalistic morphs in permanent water sites, suggesting their presence was due to factors other than faster growth allowing them to occupy ephemeral habitats. Therefore, our findings suggest environmental factors inducing the cannibalistic morphism may be more complex than previously thought.
Karraker, Nancy E.; Gibbs, James P.
It has been postulated that road deicing salts are sufficiently diluted by spring rains to ameliorate any physiological impacts to amphibians breeding in wetlands near roads. We tested this conjecture by exposing clutches of the spotted salamander (Ambystoma maculatum) to three chloride concentrations (1 mg/L, 145 mg/L, 945 mg/L) for nine days, then transferred clutches to control water for nine days, and measured change in mass at three-day intervals. We measured mass change because water uptake by clutches reduces risks to embryos associated with freezing, predation, and disease. Clutches in controls sequestered water asymptotically. Those in the moderate concentrations lost 18% mass initially and regained 14% after transfer to control water. Clutches in high concentration lost 33% mass and then lost an additional 8% after transfer. Our results suggest that spring rains do not ameliorate the effects of deicing salts in wetlands with extremely high chloride concentrations. - Road deicing salts irreversibly disrupts osmoregulation of salamander egg clutches.
Karraker, Nancy E; Gibbs, James P
It has been postulated that road deicing salts are sufficiently diluted by spring rains to ameliorate any physiological impacts to amphibians breeding in wetlands near roads. We tested this conjecture by exposing clutches of the spotted salamander (Ambystoma maculatum) to three chloride concentrations (1 mg/L, 145 mg/L, 945 mg/L) for nine days, then transferred clutches to control water for nine days, and measured change in mass at three-day intervals. We measured mass change because water uptake by clutches reduces risks to embryos associated with freezing, predation, and disease. Clutches in controls sequestered water asymptotically. Those in the moderate concentrations lost 18% mass initially and regained 14% after transfer to control water. Clutches in high concentration lost 33% mass and then lost an additional 8% after transfer. Our results suggest that spring rains do not ameliorate the effects of deicing salts in wetlands with extremely high chloride concentrations. Copyright Â© 2010 Elsevier Ltd. All rights reserved.
Karraker, Nancy E; Ruthig, Gregory R
Some causative agents of amphibian declines act synergistically to impact individual amphibians and their populations. In particular, pathogenic water molds (aquatic oomycetes) interact with environmental stressors and increase mortality in amphibian embryos. We documented colonization of eggs of three amphibian species, the wood frog (Rana sylvatica), the green frog (Rana clamitans), and the spotted salamander (Ambystoma maculatum), by water molds in the field and examined the interactive effects of road deicing salt and water molds, two known sources of mortality for amphibian embryos, on two species, R. clamitans and A. maculatum in the laboratory. We found that exposure to water molds did not affect embryonic survivorship in either A. maculatum or R. clamitans, regardless of the concentration of road salt to which their eggs were exposed. Road salt decreased survivorship of A. maculatum, but not R. clamitans, and frequency of malformations increased significantly in both species at the highest salinity concentration. The lack of an effect of water molds on survival of embryos and no interaction between road salt and water molds indicates that observations of colonization of these eggs by water molds in the field probably represent a secondary invasion of unfertilized eggs or of embryos that had died of other causes. Given increasing salinization of freshwater habitats on several continents and the global distribution of water molds, our results suggest that some amphibian species may not be susceptible to the combined effects of these factors, permitting amphibian decline researchers to devote their attention to other potential causes.
Walls, Susan C.; Barichivich, William J.; Brown, Mary E.
The Class Amphibia is one of the most severely impacted taxa in an on-going global biodiversity crisis. Because amphibian reproduction is tightly associated with the presence of water, climatic changes that affect water availability pose a particularly menacing threat to both aquatic and terrestrial-breeding amphibians. We explore the impacts that one facet of climate change—that of extreme variation in precipitation—may have on amphibians. This variation is manifested principally as increases in the incidence and severity of both drought and major storm events. We stress the need to consider not only total precipitation amounts but also the pattern and timing of rainfall events. Such rainfall “pulses” are likely to become increasingly more influential on amphibians, especially in relation to seasonal reproduction. Changes in reproductive phenology can strongly influence the outcome of competitive and predatory interactions, thus potentially altering community dynamics in assemblages of co-existing species. We present a conceptual model to illustrate possible landscape and metapopulation consequences of alternative climate change scenarios for pond-breeding amphibians, using the Mole Salamander, Ambystoma talpoideum, as an example. Although amphibians have evolved a variety of life history strategies that enable them to cope with environmental uncertainty, it is unclear whether adaptations can keep pace with the escalating rate of climate change. Climate change, especially in combination with other stressors, is a daunting challenge for the persistence of amphibians and, thus, the conservation of global biodiversity.
Collins, Sara J.; Russell, Ronald W.
The deposition of chemical pollutants into roadside wetlands from runoff is a current environmental concern. In northern latitudes, a major pollutant in runoff water is salt (NaCl), used as de-icing agents. In this study, 26 roadside ponds were surveyed for amphibian species richness and chloride concentration. Acute toxicity tests (LC 50 ) were performed on five locally common amphibian species using a range of environmentally significant NaCl concentrations. Field surveys indicated that spotted salamanders (Ambystoma maculatum) and wood frogs (Rana sylvatica) did not occupy high chloride ponds. American toads (Bufo americanus) showed no pond preference based on chloride concentration. Acute toxicity tests showed spotted salamanders and wood frogs were most sensitive to chloride, and American toads were the least. Spring peepers (Pseudacris crucifer) and green frogs (Rana clamitans) showed intermediate sensitivities. We concluded that chloride concentrations in ponds due to application of de-icing salts, influenced community structure by excluding salt intolerant species. - Salt toxicity is presented as a mechanism affecting the distribution of amphibians and structure of amphibian communities in roadside wetlands
Rowland, Freya E; Rawlings, Madelyn B; Semlitsch, Raymond D
Primary production can be controlled through bottom-up (e.g., resources) or top-down (e.g., predators) constraints. Two key bottom-up resources in small aquatic systems are light and nutrients, and forest canopy cover heavily influences these factors, whereas amphibian and invertebrate colonizers exert top-down pressure as grazers and predators. We designed our experiment to specifically manipulate two different top-down and bottom-up factors. We manipulated resources by altering light (low/high) and nutrient (low/high) availability; omnivores with the presence/absence of southern leopard frog tadpoles (Lithobates sphenocephalus); and predators with the presence/absence of spotted salamander larvae (Ambystoma maculatum) in a full-factorial experiment conducted over 14 weeks. We observed that both bottom-up and top-down effects were important in predicting lower trophic level biomass. We found a significant top-down effect of salamanders on Daphnia, but tadpoles had the strongest overall effect on the food web, influencing phytoplankton (+), periphyton (-), and chironomids (-). None of our models were good predictors of phytoplankton biomass, but both shading and nutrient availability relatively equally boosted periphyton biomass. We also found large temporal differences in food-web dynamics. Our results underscore the need for more information into how ecosystem functioning could be altered by land use, amphibian extirpation, and climate change.
Harper, Elizabeth B; Patrick, David A; Gibbs, James P
Forest loss is a primary cause of worldwide amphibian decline. Timber harvesting in the United States has caused dramatic changes in quality and extent of forest ecosystems, and intensive forest management still occurs. Although numerous studies have documented substantial reductions in amphibian densities related to timber harvest, subsequent extinctions are rare. To better understand the population dynamics that have allowed so many amphibian species to persist in the face of widespread forest disturbance, we developed spatially explicit metapopulation models for four forest-dependent amphibian species (Lithobates sylvaticus, Ambystoma opacum, A. talpoideum, and A. maculatum) that incorporated demographic and habitat selection data derived from experiments conducted as part of the Land Use Effects on Amphibian Populations Project (LEAP). We projected local and landscape-scale population persistence under 108 different forestry practice scenarios, varying treatment (partial cut, clear-cut with coarse woody debris [CWD] removed, and clearcut with CWD retained), cut patch size (1, 10, or 50 ha), total area cut (10, 20, or 30%), and initial amphibian population size (5, 50, or 500 adult females per local breeding population). Under these scenarios, landscape-scale extinction was highly unlikely, occurring in forestry practices.
Desnitskiy, Alexey G; Litvinchuk, Spartak N
The order Caudata includes about 660 species and displays a variety of important developmental traits such as cleavage pattern and egg size. However, the cleavage process of tailed amphibians has never been analyzed within a phylogenetic framework. We use published data on the embryos of 36 species concerning the character of the third cleavage furrow (latitudinal, longitudinal or variable) and the magnitude of synchronous cleavage period (up to 3-4 synchronous cell divisions in the animal hemisphere or a considerably longer series of synchronous divisions followed by midblastula transition). Several species from basal caudate families Cryptobranchidae (Andrias davidianus and Cryptobranchus alleganiensis) and Hynobiidae (Onychodactylus japonicus) as well as several representatives from derived families Plethodontidae (Desmognathus fuscus and Ensatina eschscholtzii) and Proteidae (Necturus maculosus) are characterized by longitudinal furrows of the third cleavage and the loss of synchrony as early as the 8-cell stage. By contrast, many representatives of derived families Ambystomatidae and Salamandridae have latitudinal furrows of the third cleavage and extensive period of synchronous divisions. Our analysis of these ontogenetic characters mapped onto a phylogenetic tree shows that the cleavage pattern of large, yolky eggs with short series of synchronous divisions is an ancestral trait for the tailed amphibians, while the data on the orientation of third cleavage furrows seem to be ambiguous with respect to phylogeny. Nevertheless, the midblastula transition, which is characteristic of the model species Ambystoma mexicanum (Caudata) and Xenopus laevis (Anura), might have evolved convergently in these two amphibian orders.
Taniguchi, Yuka; Kurth, Thomas; Medeiros, Daniel Meulemans; Tazaki, Akira; Ramm, Robert; Epperlein, Hans-Henning
Mesenchyme is an embryonic precursor tissue that generates a range of structures in vertebrates including cartilage, bone, muscle, kidney, and the erythropoietic system. Mesenchyme originates from both mesoderm and the neural crest, an ectodermal cell population, via an epithelial to mesenchymal transition (EMT). Because ectodermal and mesodermal mesenchyme can form in close proximity and give rise to similar derivatives, the embryonic origin of many mesenchyme-derived tissues is still unclear. Recent work using genetic lineage tracing methods have upended classical ideas about the contributions of mesodermal mesenchyme and neural crest to particular structures. Using similar strategies in the Mexican axolotl (Ambystoma mexicanum), and the South African clawed toad (Xenopus laevis), we traced the origins of fin mesenchyme and tail muscle in amphibians. Here we present evidence that fin mesenchyme and striated tail muscle in both animals are derived solely from mesoderm and not from neural crest. In the context of recent work in zebrafish, our experiments suggest that trunk neural crest cells in the last common ancestor of tetrapods and ray-finned fish lacked the ability to form ectomesenchyme and its derivatives.
Boone, M.D.; Semlitsch, R.D.; Little, E.E.; Doyle, M.C.
A leading hypothesis of amphibian population declines is that combinations of multiple stressors contribute to declines. We examined the role that chemical contamination, competition, and predation play singly and in combination in aquatic amphibian communities. We exposed larvae of American toads (Bufo americanus), southern leopard frogs (Rana sphenocephala), and spotted salamanders (Ambystoma maculatum) to overwintered bullfrog tadpoles (R. catesbeiana), bluegill sunfish (Lepomis macrochirus), the insecticide carbaryl, and ammonium nitrate fertilizer in 1000-L mesocosms. Most significantly, our study demonstrated that the presence of multiple factors reduced survival of B. americanus and A. maculatum and lengthened larval periods of R. sphenocephala. The presence of bluegill had the largest impact on the community; it eliminated B. americanus and A. maculatum and reduced the abundance of R. sphenocephala. Chemical contaminants had the second strongest effect on the community with the insecticide, reducing A. maculatum abundance by 50% and increasing the mass of anurans (frogs and toads) at metamorphosis; the fertilizer positively influenced time and mass at metamorphosis for both anurans and A. maculatum. Presence of overwintered bullfrogs reduced mass and increased time to metamorphosis of anurans. While both bluegill and overwintered bullfrog tadpoles had negative effects on the amphibian community, they performed better in the presence of one another and in contaminated habitats. Our results indicate that predicting deleterious combinations from single-factor effects may not be straightforward. Our research supports the hypothesis that combinations of factors can negatively impact some amphibian species and could contribute to population declines. ?? 2007 by the Ecological Society of America.
The interactions of GH's, PRL's and PL's with receptors for GH and PRL were examined from a comparative and evolutionary viewpoint. The binding of 125 I-bGH to membrane preparations from liver of representatives of the major classes of non-mammalian vertebrates was also studied. Only hepatic membranes from sturgeon and Gillichthys had significant bGH binding and were further characterized and compared with male rabbit liver membranes in terms of time, temperature, pH, and membrane concentration to optimize binding conditions. The binding of several members of the GH, PRL, PL family of hormones to GH receptors from liver of sturgeon, Gillichthys, rabbit, mouse and rat was investigated. in terms of hormonal specificity, the mammalian receptors and the sturgeon binding sites were similar, while Gillichthys receptors had a different pattern of hormonal specificity. The binding of 125 I-oPRL to renal membranes of the turtle, Pseudemys scripta elegans, was characterized and compared to PRL binding sites of kidney membranes of the bullfrog, Rana catesbeiana, and the tiger salamander, Ambystoma tigrinum
Diogo, R; Abdala, V; Aziz, M A; Lonergan, N; Wood, B A
In a recent study Diogo & Abdala [(2007) J Morphol 268, 504-517] reported the results of the first part of a research project on the comparative anatomy, homologies and evolution of the pectoral muscles of osteichthyans (bony fish and tetrapods). That report mainly focused on actinopterygian fish but also compared these fish with certain non-mammalian sarcopterygians. This study, which reports the second part of the research project, focuses mainly on sarcopterygians and particularly on how the pectoral and forelimb muscles have evolved during the transitions from sarcopterygian fish and non-mammalian tetrapods to monotreme and therian mammals and humans. The data obtained by our own dissections of all the pectoral and forelimb muscles of representative members of groups as diverse as sarcopterygian fish, amphibians, reptiles, monotremes and therian mammals such as rodents, tree-shrews, colugos and primates, including humans, are compared with the information available in the literature. Our observations and comparisons clearly stress that, with regard to the number of pectoral and forelimb muscles, the most striking transition within sarcopterygian evolutionary history was that leading to the origin of tetrapods. Whereas extant sarcopterygian fish have an abductor and adductor of the fin and a largely undifferentiated hypaxial and epaxial musculature, extant salamanders such as Ambystoma have more than 40 pectoral and forelimb muscles. There is no clear increase in the number of pectoral and forelimb muscles within the evolutionary transition that led to the origin of mammals and surely not to that leading to the origin of primates and humans.
Boone, Michelle D; Semlitsch, Raymond D; Mosby, Cory
Managing areas designed for human recreation so that they are compatible with natural amphibian populations can reduce the negative impacts of habitat destruction. We examined the potential for amphibians to complete larval development in golf course ponds in the presence or absence of overwintered bullfrog tadpoles (Rana catesbeiana), which are frequently found in permanent, human-made ponds. We reared larval American toads (Bufo americanus), southern leopard frogs (R. sphenocephala), and spotted salamanders (Ambystoma maculatum) with 0 or 5 overwintered bullfrog tadpoles in field enclosures located in ponds on golf courses or in experimental wetlands at a reference site. Survival to metamorphosis of American toads, southern leopard frogs, and spotted salamanders was greater in ponds on golf courses than at reference sites. We attributed this increased survival to low abundance of insect predators in golf course ponds. The presence of overwintered bullfrogs, however, reduced the survival of American toads, southern leopard frogs, and spotted salamanders reared in golf course ponds, indicating that the suitability of the aquatic habitats for these species partly depended on the biotic community present. Our results suggest that ponds in human recreational areas should be managed by maintaining intermediate hydroperiods, which will reduce the presence of bullfrog tadpoles and predators, such as fish, and which may allow native amphibian assemblages to flourish.
Busse, U; Séguin, C
To characterize molecular interactions between cells in the early amphibian embryo, we have isolated cDNAs for two members of the axolotl (Ambystoma mexicanum) Wnt family, Awnt-5A and Awnt-5B. The encoded proteins share 83% amino acid identity. Using a reverse transcription-polymerase chain reaction (RT-PCR) assay, we find that Awnt-5A transcripts are abundant in the blastula until gastrulation, barely detectable during gastrulation, and increase again during neurulation. They are detected throughout the remaining development and in hatched larvae. In contrast, transcripts for Awnt-5B are undetectable in the blastula. They appear with gastrulation, are present throughout neurulation and organogenesis, and decrease to barely detectable levels in hatched larvae. PCR reactions performed using cDNA library-phage DNA templates derived from whole neurulae versus embryos with the neuroectoderm removed suggest that, in the neurula, Awnt-5A transcripts are present in neuroectodermal as well as non-neuroectodermal tissues while Awnt-5B mRNAs are predominantly localized in the neuroectoderm. To localize Awnt-5A expression in embryos before gastrulation, early gastrulae were dissected by cutting along the animal-vegetal and future dorso-ventral axes and analyzed by RT-PCR. At this early stage, Awnt-5A transcripts appear to be predominantly localized in the dorso-vegetal region of the embryo. These results suggest that the two closely related Awnt-5 genes participate in different morphogenetic processes during early axolotl development.
Wischin, Sabina; Castañeda-Patlán, Cristina; Robles-Flores, Martha; Chimal-Monroy, Jesús
Limb regeneration involves several interrelated physiological processes in which a particular signalling pathway may play a variety of functions. Blocking the function of Wnt/β-catenin signalling during limb regeneration inhibits regeneration in axolotls (Ambystoma mexicanum). Limb development shares many features with limb regeneration, and Wnt/β-catenin activation has different effects depending on the developmental stage. The aim of this study was to evaluate whether Wnt/β-catenin signalling activation during axolotl limb regeneration has different effects when activated at different stages of regeneration. To evaluate this hypothesis, we treated amputated axolotls with a Wnt agonist chemical at different stages of limb regeneration. The results showed that limb regeneration was inhibited when the treatment began before blastema formation. Under these conditions, blastema formation was hindered, possibly due to the lack of innervation. On the other hand, when axolotls were treated after blastema formation and immediately before the onset of morphogenesis, we observed structural disorganization in skeletal formation. In conclusion, we found that limb regeneration was differentially affected depending on the stage at which the Wnt signalling pathway was activated. Copyright © 2017 Elsevier B.V. All rights reserved.
Jeremiah R. Alexander
Full Text Available Understanding the temporal and spatial scale at which habitat alteration impacts populations is important for conservation and management. Amphibians have declined more than other vertebrates, and pond-breeding species are particularly susceptible to habitat loss and fragmentation because they have terrestrial and aquatic life stages. One approach to management of pond-breeding species is protection of core upland habitat surrounding the breeding pond. We used genetic variation as an indicator of population status in a common amphibian species, spotted salamanders (Ambystoma maculatum, to determine how amount of suitable upland habitat relates to population status in the greater Charlotte, North Carolina, USA metropolitan area. We developed candidate models to evaluate the relative influence of historical and contemporary forested habitat availability on population genetic variation at two spatial scales of upland area (164 m and 2000 m at four time intervals over the past seven decades (1938, 1978, 1993, 2005. We found that historical land cover best predicted contemporary allelic richness. Inbreeding coefficient and observed heterozygosity were not effectively predicted by forest cover at either spatial or temporal scales. Allelic richness was best predicted at the smaller spatial scale in the 1993 time interval. Predicting and understanding how future landscape configuration affects genetic variation of common and rare species is imperative for the conservation of amphibian and other wildlife populations.
Chen, Xiaoping; Song, Fengyu; Jhamb, Deepali; Li, Jiliang; Bottino, Marco C.; Palakal, Mathew J.; Stocum, David L.
We tested the ability of the axolotl (Ambystoma mexicanum) fibula to regenerate across segment defects of different size in the absence of intervention or after implant of a unique 8-braid pig small intestine submucosa (SIS) scaffold, with or without incorporated growth factor combinations or tissue protein extract. Fractures and defects of 10% and 20% of the total limb length regenerated well without any intervention, but 40% and 50% defects failed to regenerate after either simple removal of bone or implanting SIS scaffold alone. By contrast, scaffold soaked in the growth factor combination BMP-4/HGF or in protein extract of intact limb tissue promoted partial or extensive induction of cartilage and bone across 50% segment defects in 30%-33% of cases. These results show that BMP-4/HGF and intact tissue protein extract can promote the events required to induce cartilage and bone formation across a segment defect larger than critical size and that the long bones of axolotl limbs are an inexpensive model to screen soluble factors and natural and synthetic scaffolds for their efficacy in stimulating this process. PMID:26098852
Wang, Supen; Liu, Conghui; Wilson, Anthony B; Zhao, Na; Li, Xianping; Zhu, Wei; Gao, Xu; Liu, Xuan; Li, Yiming
The identification of the factors responsible for genetic variation and differentiation at adaptive loci can provide important insights into the evolutionary process and is crucial for the effective management of threatened species. We studied the impact of environmental viral richness and abundance on functional diversity and differentiation of the MHC class Ia locus in populations of the black-spotted pond frog (Pelophylax nigromaculatus), an IUCN-listed species, on 24 land-bridge islands of the Zhoushan Archipelago and three nearby mainland sites. We found a high proportion of private MHC alleles in mainland and insular populations, corresponding to 32 distinct functional supertypes, and strong positive selection on MHC antigen-binding sites in all populations. Viral pathogen diversity and abundance were reduced at island sites relative to the mainland, and islands housed distinctive viral communities. Standardized MHC diversity at island sites exceeded that found at neutral microsatellites, and the representation of key functional supertypes was positively correlated with the abundance of specific viruses in the environment (Frog virus 3 and Ambystoma tigrinum virus). These results indicate that pathogen-driven diversifying selection can play an important role in maintaining functionally important MHC variation following island isolation, highlighting the importance of considering functionally important genetic variation and host-pathogen associations in conservation planning and management. © 2017 John Wiley & Sons Ltd.
Anatomy of the pectoral and forelimb muscles of wildtype and green fluorescent protein-transgenic axolotls and comparison with other tetrapods including humans: a basis for regenerative, evolutionary and developmental studies.
Diogo, R; Tanaka, E M
The axolotl Ambystoma mexicanum is one of the most used model organisms in evolutionary, developmental and regenerative studies, particularly because it can reconstitute a fully functional and complete forelimb/hindlimb. Surprisingly, there is no publication that describes all the pectoral and forelimb muscles of this species or provides a comparative framework between these muscles and those of other model organisms and of modern humans. In the present paper we describe and illustrate all these muscles in A. mexicanum and provide the first report about the myology of adults of a model organism that is based on analyses and dissections of both wildtype animals and transgenic animals that express green fluorescent protein (GFP) in muscle fibers. On the one hand, the inclusion of GFP-transgenic animals allows us to show the muscles as more commonly seen, and thus easier to understand, by current developmental and regenerative biologists. On the other hand, by including wildtype and GFP-transgenic animals and by visualizing these latter animals with and without a simultaneous transmission laser light, we were able to obtain a more complete and clearer understanding of the exact limit of the fleshy and tendinous parts of the muscles and their specific connections with the skeletal elements. This in turn allowed us to settle some controversies in previous anatomical and comparative studies. As most developmental, regenerative and evolutionary biologists are interested in comparing their observations of A. mexicanum with observations in other model organisms, and ultimately in using this information to increase the understanding of human evolution and medicine, we also provide tables showing the homologies between the pectoral and forelimb muscles of axolotls, of model organisms such as mice, frogs and chicken, and of Homo sapiens. An example illustrating the outcomes of using our methodology and of our observations is that they revealed that, contrary to what is often
Nicholson, Abbigail E; Siegel, Dustin S
Male salamanders use nephrons from the genital kidney to transport sperm from the testicular lobules to the Wolffian duct. The microstructure of the epithelia of the genital kidney proximal tubule and distal tubule was studied over 1 year in a population of Notophthalmus viridescens from Crawford and Pike counties in central Missouri. Through ultrastructural analysis, we were able to support the hypothesis that the genital kidney nephrons are modified to aid in the transportation of sperm. A lack of folding of the basal plasma membrane, in both the genital kidney proximal and distal tubules when compared to the pelvic kidney proximal and distal tubules, reduces the surface area and thus likely decreases the efficiency of reabsorption in these nephron regions of the genital kidney. Ciliated epithelial cells are also present along the entire length of the genital kidney proximal tubule, but are lacking in the epithelium of the pelvic kidney proximal tubule. The exact function of these cilia remains unknown, but they may aid in mixing of seminal fluids or the transportation of immature sperm through the genital kidney nephrons. Ultrastructural analysis of proximal and distal tubules of the genital kidney revealed no seasonal variation in cellular activity and no mass production of seminal fluids throughout the reproductive cycle. Thus, we failed to support the hypothesis that the cellular activity of the epithelia lining the genital kidney nephrons is correlated to specific events in the reproductive cycle. The cytoplasmic contents and overall structure of the genital and pelvic kidney epithelial cells were similar to recent observations in Ambystoma maculatum, with the absence of abundant dense bodies apically in the epithelial cells lining the genital kidney distal tubule. © 2014 Wiley Periodicals, Inc.
Kawano, Sandy M; Blob, Richard W
The invasion of land was a pivotal event in vertebrate evolution that was associated with major appendicular modifications. Although fossils indicate that the evolution of fundamentally limb-like appendages likely occurred in aquatic environments, the functional consequences of using early digited limbs, rather than fins, for terrestrial propulsion have had little empirical investigation. Paleontological and experimental analyses both have led to the proposal of an early origin of "hind limb-driven" locomotion among tetrapods or their ancestors. However, the retention of a pectoral appendage that had already developed terrestrial adaptations has been proposed for some taxa, and few data are available from extant functional models that can provide a foundation for evaluating the relative contributions of pectoral and pelvic appendages to terrestrial support among early stem tetrapods. To examine these aspects of vertebrate locomotor evolution during the invasion of land, we measured three-dimensional ground reaction forces (GRFs) produced by isolated pectoral fins of mudskipper fishes (Periophthalmus barbarus) during terrestrial crutching, and compared these to isolated walking footfalls by the forelimbs and hind limbs of tiger salamanders (Ambystoma tigrinum), a species with subequally-sized limbs that facilitate comparisons to early tetrapods. Pectoral appendages of salamanders and mudskippers exhibited numerous differences in GRFs. Compared with salamander forelimbs, isolated fins of mudskippers bear lower vertical magnitudes of GRFs (as a proportion of body weight), and had GRFs that were oriented more medially. Comparing the salamanders' forelimbs and hind limbs, although the peak net GRF occurs later in stance for the forelimb, both limbs experience nearly identical mediolateral and vertical components of GRF, suggesting comparable contributions to support. Thus, forelimbs could also have played a significant locomotor role among basal tetrapods that had limbs
Niemiller, M.L.; Graham, Reynolds R.; Glorioso, B.M.; Spiess, J.; Miller, B.T.
The cedar glades and barrens of the Inner Central Basin (ICB) of middle Tennessee support a unique and diverse flora and fauna and represent some of the state's most valued natural areas. We conducted herpetofaunal inventories of the cedar glades, associated barrens, cedar-hardwood forest, and adjacent aquatic habitats of the Stones River drainage of Middle Tennessee, focusing our sampling effort primarily at seven state- or federally owned properties in Rutherford and Wilson counties. These properties included Stones River National Battlefield (SRNB), Flat Rock State Natural Area (FRSNA), Vesta Cedar Glade State Natural Area (VSNA), Fall Creek Recreation Area (FCRA) on J. Percy Priest Wildlife Management Area, Cedars of Lebanon State Forest (CLSF), Cedars of Lebanon State Forest Natural Area (CLSNA), and Cedars of Lebanon State Park (CLSP). We used a variety of inventory techniques in terrestrial, aquatic, and subterranean habitats to survey these properties periodically from 1989 to 2010. We documented 49 species (22 amphibian and 27 reptile) accounting for 75.4% of the 65 herpetofaunal species thought to occur in the ICB, including records for Cemophora coccinea, Aneides aeneus, Gyrinophilus palleucus, Ambystoma barbouri, and Pseudotriton montanus. We found differences in alpha and beta diversity between sites, with the CLSF complex containing a high of 41 herpetofaunal species and FRSNA containing a low of 23 species. Beta diversity comparisons indicated similarity in amphibian species composition between FRSNA and CLSF and between SRNB and CLSF (9 shared species), and in reptile species composition between VSNA and the CLSF complex (16 shared species). We compare the results of our inventory with two previous studies conducted in the area and discuss the relative abundance, conservation, and threats to the herpetofaunal community of these habitats.
Direnberger, Stephan; Banchi, Roberto; Brosel, Sonja; Seebacher, Christian; Laimgruber, Stefan; Uhl, Rainer; Felmy, Felix; Straka, Hans; Kunz, Lars
Optical visualization of neural network activity is limited by imaging system-dependent technical tradeoffs. To overcome these constraints, we have developed a powerful low-cost and flexible imaging system with high spectral variability and unique spatio-temporal precision for simultaneous optical recording and manipulation of neural activity of large cell groups. The system comprises eight high-power light-emitting diodes, a camera with a large metal-oxide-semiconductor sensor and a high numerical aperture water-dipping objective. It allows fast and precise control of excitation and simultaneous low noise imaging at high resolution. Adjustable apertures generated two independent areas of variable size and position for simultaneous optical activation and image capture. The experimental applicability of this system was explored in semi-isolated preparations of larval axolotl (Ambystoma mexicanum) with intact inner ear organs and central nervous circuits. Cyclic galvanic stimulation of semicircular canals together with glutamate- and γ-aminobutyric acid (GABA)-uncaging caused a corresponding modulation of Ca(2+) transients in central vestibular neurons. These experiments revealed specific cellular properties as well as synaptic interactions between excitatory and inhibitory inputs, responsible for spatio-temporal-specific sensory signal processing. Location-specific GABA-uncaging revealed a potent inhibitory shunt of vestibular nerve afferent input in the predominating population of tonic vestibular neurons, indicating a considerable impact of local and commissural inhibitory circuits on the processing of head/body motion-related signals. The discovery of these previously unknown properties of vestibular computations demonstrates the merits of our novel microscope system for experimental applications in the field of neurobiology. © 2015 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.
Full Text Available Although in recent years the study of gene expression variation in the absence of genetic or environmental cues or gene expression heterogeneity has intensified considerably, many basic and applied biological fields still remain unaware of how useful the study of gene expression heterogeneity patterns might be for the characterization of biological systems and/or processes. Largely based on the modulator effect chromatin compaction has for gene expression heterogeneity and the extensive changes in chromatin compaction known to occur for specialized cells that are naturally or artificially induced to revert to less specialized states or dedifferentiate, I recently hypothesized that processes that concur with cell dedifferentiation would show an extensive reduction in gene expression heterogeneity. The confirmation of the existence of such trend could be of wide interest because of the biomedical and biotechnological relevance of cell dedifferentiation-based processes, i.e., regenerative development, cancer, human induced pluripotent stem cells, or plant somatic embryogenesis. Here, I report the first empirical evidence consistent with the existence of an extensive reduction in gene expression heterogeneity for processes that concur with cell dedifferentiation by analyzing transcriptome dynamics along forearm regenerative development in Ambystoma mexicanum or axolotl. Also, I briefly discuss on the utility of the study of gene expression heterogeneity dynamics might have for the characterization of cell dedifferentiation-based processes, and the engineering of tools that afforded better monitoring and modulating such processes. Finally, I reflect on how a transitional reduction in gene expression heterogeneity for dedifferentiated cells can promote a long-term increase in phenotypic heterogeneity following cell dedifferentiation with potential adverse effects for biomedical and biotechnological applications.
Jonathan P Rose
Full Text Available Species distribution models (SDMs are increasingly used to project the potential distribution of introduced species outside their native range. Such studies rarely explicitly evaluate potential conflicts with native species should the range of introduced species expand. Two snake species native to eastern North America, Nerodia fasciata and Nerodia sipedon, have been introduced to California where they represent a new stressor to declining native amphibians, fish, and reptiles. To project the potential distributions of these non-native watersnakes in western North America, we built ensemble SDMs using MaxEnt, Boosted Regression Trees, and Random Forests and habitat and climatic variables. We then compared the overlap between the projected distribution of invasive watersnakes and the distributions of imperiled native amphibians, fish, and reptiles that can serve as prey or competitors for the invaders, to estimate the risk to native species posed by non-native watersnakes. Large areas of western North America were projected to be climatically suitable for both species of Nerodia according to our ensemble SDMs, including much of central California. The potential distributions of both N. fasciata and N. sipedon overlap extensively with the federally threatened Giant Gartersnake, Thamnophis gigas, which inhabits a similar ecological niche. N. fasciata also poses risk to the federally threatened California Tiger Salamander, Ambystoma californiense, whereas N. sipedon poses risk to some amphibians of conservation concern, including the Foothill Yellow-legged Frog, Rana boylii. We conclude that non-native watersnakes in California can likely inhabit ranges of several native species of conservation concern that are expected to suffer as prey or competing species for these invaders. Action should be taken now to eradicate or control these invasions before detrimental impacts on native species are widespread. Our methods can be applied broadly to quantify
Karraker, Nancy E; Gibbs, James P; Vonesh, James R
Deicing agents, primarily road salt, are applied to roads in 26 states in the United States and in a number of European countries, yet the scale of impacts of road salt on aquatic organisms remains largely under-studied. The issue is germane to amphibian conservation because both adult and larval amphibians are known to be particularly sensitive to changes in their osmolar environments. In this study, we combined survey, experimental, and demographic modeling approaches to evaluate the possible effects of road salt on two common vernal-pond-breeding amphibian species, the spotted salamander (Ambystoma maculatum) and the wood frog (Rana sylvatica). We found that in the Adirondack Mountain Region of New York (USA), road salt traveled up to 172 m from the highway into wetlands. Surveys showed that egg mass densities of spotted salamanders (A. maculatum) and wood frogs (R. sylvatica) were two times higher in forest pools than roadside pools, but this pattern was better explained by road proximity than by increased salinity. Experiments demonstrated that embryonic and larval survival were reduced at moderate (500 muS) and high conductivities (3000 muS) in A. maculatum and at high conductivities in R. sylvatica. Demographic models suggest that such egg and larval stage effects of salt may have important impacts on populations near roads, particularly in the case of A. maculatum, for which salt exposure may lead to local extinction. For both species, the effect of road salt was dependent upon the strength of larval density dependence and declined rapidly with distance from the roadside, with the greatest negative effects being limited to within 50 m. Based on this evidence, we argue that efforts to protect local populations of A. maculatum and R. sylvatica in roadside wetlands should, in part, be aimed at reducing application of road salt near wetlands with high conductivity levels.
Chandler, C H; Zamudio, K R
The outcome of sexual selection on males may depend on female mate choice and male-male competition as well as postcopulatory processes such as cryptic female choice and sperm competition. We studied the outcome of sexual selection in the spotted salamander (Ambystoma maculatum), specifically examining the role of body size and relatedness on male reproductive success. Using controlled mating experiments in the field, we gave females access to three males of different sizes. We used seven microsatellite loci to determine paternity in the resulting larvae, estimate relatedness (r) between females and their mates, and calculate md(2) (a measure of within-individual genomic divergence), heterozygosity, and standardized heterozygosity in the larvae. Both body size and relatedness to the female were significant predictors of male reproductive success. The relatedness of the males available to a female did not influence the amount of stored sperm she used to sire her larvae. Nonetheless, computer simulations showed that the average md(2), heterozygosity, and standardized heterozygosity of the offspring were lower than expected by random mating. These differences are due to the use of stored sperm to fertilize some eggs; md(2), heterozygosity, and standardized heterozygosity of larvae sired by stored sperm were significantly lower than those of larvae sired by the experimental males. These results suggest that relatedness may further influence a male's long-term reproductive success by determining whether his sperm is stored for later breeding seasons. Sexual selection in this salamander likely involves a complex interaction among many factors and may act over many seasons.
Boone, M.D.; James, S.M.
Amphibians developing in wetlands embedded within or near agricultural lands may frequently encounter chemical mixtures. The objectives of our study were to determine the effects that post-application concentrations of an insecticide (carbaryl) and an herbicide (atrazine) have on body mass, development, and survival of two anuran species (southern leopard frog, Rana sphenocephala; American toad, Bufo americanus) and two caudate species (spotted salamander, Ambystoma maculatum; small-mouthed salamander, A. texanum) reared in outdoor cattle tank mesocosms. In one experiment, we manipulated tadpole density (low or high), carbaryl exposure (0, 3.5, 7.0 mg/L), and atrazine exposure (0 or 200 μg/L) to test for effects on development, mass, and survival of larvae. In a second experiment, we manipulated pond hydroperiod (constant or drying), carbaryl exposure (0 or 5 mg/L), and atrazine exposure (0 or 200 μg/L) to test for effects on mass, time, and survival to metamorphosis. Salamanders were virtually eliminated in carbaryl treatments, indicating that at realistic levels, this insecticide could cause population declines for salamanders in contaminated habitats. Carbaryl also had negative effects on toad survival. Exposure to atrazine had negative effects on body size, development, and time to metamorphosis in anuran species, which were associated with reduced chlorophyll levels. Both chemicals interacted significantly with density or hydroperiod, indicating that the environmental conditions could influence the impact of a contaminant. A significant atrazine-by-carbaryl interaction resulted in smaller and less developed spotted salamander larvae than in control ponds. Atrazine exposure, however, appeared to moderate negative effects of carbaryl for spotted salamanders. Our research suggests that important changes in the community's food web result from chemical exposure, which influence the susceptibility of amphibian species to contaminants.
Rohr, Jason R; Palmer, Brent D
Climate change is believed to be causing declines of ectothermic vertebrates, but there is little evidence that climatic conditions associated with declines have exceeded critical (i.e., acutely lethal) maxima or minima, and most relevant studies are correlative, anecdotal, or short-term (hours). We conducted an 11-week factorial experiment to examine the effects of temperature (22 °C or 27 °C), moisture (wet or dry), and atrazine (an herbicide; 0, 4, 40, 400 μg/L exposure as embryos and larvae) on the survival, growth, behavior, and foraging rates of postmetamorphic streamside salamanders (Ambystoma barbouri), a species of conservation concern. The tested climatic conditions were between the critical maxima and minima of streamside salamanders; thus, this experiment quantified the long-term effects of climate change within the noncritical range of this species. Despite a suite of behavioral adaptations to warm and dry conditions (e.g., burrowing, refuge use, huddling with conspecifics, and a reduction in activity), streamside salamanders exhibited significant loss of mass and significant mortality in all but the cool and moist conditions, which were closest to the climatic conditions in which they are most active in nature. A temperature of 27 °C represented a greater mortality risk than dry conditions; death occurred rapidly at this temperature and more gradually under cool and dry conditions. Foraging decreased under dry conditions, which suggests there were opportunity costs to water conservation. Exposure to the herbicide atrazine additively decreased water-conserving behaviors, foraging efficiency, mass, and time to death. Hence, the hypothesis that moderate climate change can cause population declines is even more plausible under scenarios with multiple stressors. These results suggest that climate change within the noncritical range of species and pollution may reduce individual performance by altering metabolic demands, hydration, and foraging effort
Possible impacts of an 800-MW coal-fired power plant to be built near Atikokan, Ontario were evaluated. It is feared that the emissions of SO/sub 2/ will lead to the deposition of sulfuric acid and result in the acidification of freshwaters in nearby parks and wilderness areas. The most obvious biological effects of acidification are damages to populations of fish. Less conspicuous but no less severe damages also occur to other organisms. It appears that all trophic levels are affected: species numbers are reduced, biomasses are altered, and primary production and decomposition are impaired. Field experiments and laboratory experiments indicate that microbial activity is reduced and that the recycling of materials is greatly impeded at low pH. This may interfere with nutrient supplies to plants and decrease the microbial biomass available to higher trophic levels. Phytoplankton densities decrease in acidified lakes and there is a reduction in some species of macrophytes. On the other hand, Sphagnum and benthic filamentous algae greatly increase in acidified conditions. The total primary productivity of lakes and streams may actually increase because of such dense growths on the bottom. Zooplankton and benthic invertebrate communities become less complex as acidity increases. This may in part be due to reduced food supplies, but direct inhibition by H/sub 2/SO/sub 4/ has also been demonstrated. This removal of fish food organisms may exacerbate damage to fisheries, especially in the pH range of 5 to 6. When a lake loses all fish because of low pH, a few species of invertebrates may become very abundant. The salamanders Ambystoma jeffersonium and A. maculatum, sensitive to acidity below pH 7.0 and 5.0 respectively, are being eliminated from small ponds or temporary pools in the region around Ithaca, NY because of the impact of acid precipitation. Species of frogs in some lakes are also being eliminated because of acidification. (ERB)
Doyle, Jacqueline M; McCormick, Cory R; DeWoody, J Andrew
Many animals, such as crustaceans, insects, and salamanders, package their sperm into spermatophores, and the number of spermatozoa contained in a spermatophore is relevant to studies of sexual selection and sperm competition. We used two molecular methods, real-time quantitative polymerase chain reaction (RT-qPCR) and spectrophotometry, to estimate sperm numbers from spermatophores. First, we designed gene-specific primers that produced a single amplicon in four species of ambystomatid salamanders. A standard curve generated from cloned amplicons revealed a strong positive relationship between template DNA quantity and cycle threshold, suggesting that RT-qPCR could be used to quantify sperm in a given sample. We then extracted DNA from multiple Ambystoma maculatum spermatophores, performed RT-qPCR on each sample, and estimated template copy numbers (i.e. sperm number) using the standard curve. Second, we used spectrophotometry to determine the number of sperm per spermatophore by measuring DNA concentration relative to the genome size. We documented a significant positive relationship between the estimates of sperm number based on RT-qPCR and those based on spectrophotometry. When these molecular estimates were compared to spermatophore cap size, which in principle could predict the number of sperm contained in the spermatophore, we also found a significant positive relationship between sperm number and spermatophore cap size. This linear model allows estimates of sperm number strictly from cap size, an approach which could greatly simplify the estimation of sperm number in future studies. These methods may help explain variation in fertilization success where sperm competition is mediated by sperm quantity. © 2010 Blackwell Publishing Ltd.
Pilliod, David S.; Arkle, Robert S.; Maxell, Bryce A.
Studies have demonstrated negative effects of non-native, predatory fishes on native amphibians, yet it is still unclear why some amphibian populations persist, while others are extirpated, following fish invasion. We examined this question by developing habitat-based occupancy models for the long-toed salamander (Ambystoma macrodactylum) and nonnative fish using survey data from 1,749 water bodies across 470 catchments in the Northern Rocky Mountains, USA. We first modeled the habitat associations of salamanders at 468 fishless water bodies in 154 catchments where non-native fish were historically, and are currently, absent from the entire catchment. Wethen applied this habitat model to the complete data set to predict the probability of salamander occupancy in each water body, removing any effect of fish presence. Finally, we compared field-observed occurrences of salamanders and fish to modeled probability of salamander occupancy. Suitability models indicated that fish and salamanders had similar habitat preferences, possibly resulting in extirpations of salamander populations from entire catchments where suitable habitats were limiting. Salamanders coexisted with non-native fish in some catchments by using marginal quality, isolated (no inlet or outlet) habitats that remained fishless. They rarely coexisted with fish within individual water bodies and only where habitat quality was highest. Connectivity of water bodies via streams resulted in increased probability of fish invasion and consequently reduced probability of salamander occupancy.These results could be used to identify and prioritize catchments and water bodies where control measures would be most effective at restoring amphibian populations. Our approach could be useful as a framework for improved investigations into questions of persistence and extirpation of native species when non-native species have already become established.
Miller, David A.W.; Grant, Evan H. Campbell
Regional monitoring strategies frequently employ a nested sampling design where a finite set of study areas from throughout a region are selected within which intensive sub-sampling occurs. This sampling protocol naturally lends itself to a hierarchical analysis to account for dependence among sub-samples. Implementing such an analysis within a classic likelihood framework is computationally prohibitive with species occurrence data when accounting for detection probabilities. Bayesian methods offer an alternative framework to make this analysis feasible. We demonstrate a general approach for estimating occupancy when data come from a nested sampling design. Using data from a regional monitoring program of wood frogs (Lithobates sylvaticus) and spotted salamanders (Ambystoma maculatum) in vernal pools, we analyzed data using static and dynamic occupancy frameworks. We analyzed observations from 2004-2013collected within 14 protected areas located throughout the northeast United States . We use the data set to estimate trends in occupancy at both the regional and individual protected area level. We show that occupancy at the regional level was relatively stable for both species. Much more variation occurred within individual study areas, with some populations declining and some increasing for both species. We found some evidence for a latitudinal gradient in trends among protected areas. However, support for this pattern is overestimated when the hierarchical nature of the data collection is not controlled for in the analysis. For both species, occupancy appeared to be declining in the most southern areas, while occupancy was stable or increasing in more northern areas. These results shed light on the range-level population status of these pond-breeding amphibians and our approach provides a framework that can be used to examine drivers of change including among-year and among-site variation in occurrence dynamics, while properly accounting for nested structure of
Kim, Eunsoo; Lin, Yuan; Kerney, Ryan; Blumenberg, Lili; Bishop, Cory
Egg masses of the yellow-spotted salamander Ambystoma maculatum form an association with the green alga "Oophila amblystomatis" (Lambert ex Wille), which, in addition to growing within individual egg capsules, has recently been reported to invade embryonic tissues and cells. The binomial O. amblystomatis refers to the algae that occur in A. maculatum egg capsules, but it is unknown whether this population of symbionts constitutes one or several different algal taxa. Moreover, it is unknown whether egg masses across the geographic range of A. maculatum, or other amphibians, associate with one or multiple algal taxa. To address these questions, we conducted a phylogeographic study of algae sampled from egg capsules of A. maculatum, its allopatric congener A. gracile, and two frogs: Lithobates sylvatica and L. aurora. All of these North American amphibians form associations with algae in their egg capsules. We sampled algae from egg capsules of these four amphibians from localities across North America, established representative algal cultures, and amplified and sequenced a region of 18S rDNA for phylogenetic analysis. Our combined analysis shows that symbiotic algae found in egg masses of four North American amphibians are closely related to each other, and form a well-supported clade that also contains three strains of free-living chlamydomonads. We designate this group as the 'Oophila' clade, within which the symbiotic algae are further divided into four distinct subclades. Phylogenies of the host amphibians and their algal symbionts are only partially congruent, suggesting that host-switching and co-speciation both play roles in their associations. We also established conditions for isolating and rearing algal symbionts from amphibian egg capsules, which should facilitate further study of these egg mass specialist algae.
Archer, Grace A; Phillips, Christopher A; Adamovicz, Laura; Band, Mark; Byrd, John; Allender, Matthew C
Conservation efforts are investigating the impact of diseases within a species of interest, including prevalence and transmission and morbidity and mortality rates. However, the majority of these studies focus solely on the characteristics of a single pathogen. Recently, the role of copathogens has been reported to impact disease susceptibility and mortality. To that effect, a survey was conducted including 318 eastern box turtles ( Terrapene carolina carolina) from populations in Illinois and Tennessee in 2014 and 2015. Blood samples and oral swabs were collected for quantitative polymerase chain reaction (qPCR) of 15 different pathogens performed in a multiplex format using Fluidigm array technology. Four pathogens were found with varying qPCR prevalence: ranavirus (FV3; n = 2, 0.6%), Terrapene herpesvirus 1 (TerHV1; n = 129, 40.7%), box turtle Mycoplasma sp. (BT Myco; n = 14, 4.6%), and box turtle adenovirus (BT Adv1; n = 18, 11%). Thirteen pathogens were not identified in any sample, including Mycoplasma agassizii, M. testudineum, Salmonella enteriditis, S. typhmirium, Borrelia burgdorferi, Anaplasma phagocyophilum, tortoise intranuclear coccidia, Ambystoma tigrinum virus, Bohle iridovirus, Epizootic hematopoietic necrosis virus, and testudinid herpesvirus 2. Copathogen occurrence was rare but was observed in eight individuals with TerHV1-BT Myco detection and two animals with TerHV1-Adv1. Significant differences were observed in pathogen detection across season (TerHV1, BT Adv1, BT Myco, and TerHV1-Myco) and year (TerHV1, BT Adv1, and TerHV1-Myco). The results of this survey highlight that a single pathogen model may not adequately explain pathogen dynamics and that conservation efforts need to be aimed at detecting multiple pathogens in order to fully characterize population health.
Diogo, R; Abdala, V; Aziz, M A; Lonergan, N; Wood, B A
In a recent study Diogo & Abdala [(2007) JMorphol268, 504–517] reported the results of the first part of a research project on the comparative anatomy, homologies and evolution of the pectoral muscles of osteichthyans (bony fish and tetrapods). That report mainly focused on actinopterygian fish but also compared these fish with certain non-mammalian sarcopterygians. This study, which reports the second part of the research project, focuses mainly on sarcopterygians and particularly on how the pectoral and forelimb muscles have evolved during the transitions from sarcopterygian fish and non-mammalian tetrapods to monotreme and therian mammals and humans. The data obtained by our own dissections of all the pectoral and forelimb muscles of representative members of groups as diverse as sarcopterygian fish, amphibians, reptiles, monotremes and therian mammals such as rodents, tree-shrews, colugos and primates, including humans, are compared with the information available in the literature. Our observations and comparisons clearly stress that, with regard to the number of pectoral and forelimb muscles, the most striking transition within sarcopterygian evolutionary history was that leading to the origin of tetrapods. Whereas extant sarcopterygian fish have an abductor and adductor of the fin and a largely undifferentiated hypaxial and epaxial musculature, extant salamanders such as Ambystoma have more than 40 pectoral and forelimb muscles. There is no clear increase in the number of pectoral and forelimb muscles within the evolutionary transition that led to the origin of mammals and surely not to that leading to the origin of primates and humans. PMID:19438764
Full Text Available Egg masses of the yellow-spotted salamander Ambystoma maculatum form an association with the green alga "Oophila amblystomatis" (Lambert ex Wille, which, in addition to growing within individual egg capsules, has recently been reported to invade embryonic tissues and cells. The binomial O. amblystomatis refers to the algae that occur in A. maculatum egg capsules, but it is unknown whether this population of symbionts constitutes one or several different algal taxa. Moreover, it is unknown whether egg masses across the geographic range of A. maculatum, or other amphibians, associate with one or multiple algal taxa. To address these questions, we conducted a phylogeographic study of algae sampled from egg capsules of A. maculatum, its allopatric congener A. gracile, and two frogs: Lithobates sylvatica and L. aurora. All of these North American amphibians form associations with algae in their egg capsules. We sampled algae from egg capsules of these four amphibians from localities across North America, established representative algal cultures, and amplified and sequenced a region of 18S rDNA for phylogenetic analysis. Our combined analysis shows that symbiotic algae found in egg masses of four North American amphibians are closely related to each other, and form a well-supported clade that also contains three strains of free-living chlamydomonads. We designate this group as the 'Oophila' clade, within which the symbiotic algae are further divided into four distinct subclades. Phylogenies of the host amphibians and their algal symbionts are only partially congruent, suggesting that host-switching and co-speciation both play roles in their associations. We also established conditions for isolating and rearing algal symbionts from amphibian egg capsules, which should facilitate further study of these egg mass specialist algae.
Prehn Lea R
Full Text Available Abstract Background Three kidney systems appear during vertebrate development: the pronephroi, mesonephroi and metanephroi. The pronephric duct is the first or primary ureter of these kidney systems. Its role as a key player in the induction of nephrogenic mesenchyme is well established. Here we investigate whether the duct is involved in urine modification using larvae of the freshwater amphibian Ambystoma mexicanum (axolotl as model. Results We investigated structural as well as physiological properties of the pronephric duct. The key elements of our methodology were: using histology, light and transmission electron microscopy as well as confocal laser scanning microscopy on fixed tissue and applying the microperfusion technique on isolated pronephric ducts in combination with single cell microelectrode impalements. Our data show that the fully differentiated pronephric duct is composed of a single layered epithelium consisting of one cell type comparable to the principal cell of the renal collecting duct system. The cells are characterized by a prominent basolateral labyrinth and a relatively smooth apical surface with one central cilium. Cellular impalements demonstrate the presence of apical Na+ and K+ conductances, as well as a large K+ conductance in the basolateral cell membrane. Immunolabeling experiments indicate heavy expression of Na+/K+-ATPase in the basolateral labyrinth. Conclusions We propose that the pronephric duct is important for the subsequent modification of urine produced by the pronephros. Our results indicate that it reabsorbs sodium and secretes potassium via channels present in the apical cell membrane with the driving force for ion movement provided by the Na+/K+ pump. This is to our knowledge the first characterization of the pronephric duct, the precursor of the collecting duct system, which provides a model of cell structure and basic mechanisms for ion transport. Such information may be important in understanding
Peterman, William; Brocato, Emily R; Semlitsch, Raymond D; Eggert, Lori S
In population or landscape genetics studies, an unbiased sampling scheme is essential for generating accurate results, but logistics may lead to deviations from the sample design. Such deviations may come in the form of sampling multiple life stages. Presently, it is largely unknown what effect sampling different life stages can have on population or landscape genetic inference, or how mixing life stages can affect the parameters being measured. Additionally, the removal of siblings from a data set is considered best-practice, but direct comparisons of inferences made with and without siblings are limited. In this study, we sampled embryos, larvae, and adult Ambystoma maculatum from five ponds in Missouri, and analyzed them at 15 microsatellite loci. We calculated allelic richness, heterozygosity and effective population sizes for each life stage at each pond and tested for genetic differentiation (F ST and D C ) and isolation-by-distance (IBD) among ponds. We tested for differences in each of these measures between life stages, and in a pooled population of all life stages. All calculations were done with and without sibling pairs to assess the effect of sibling removal. We also assessed the effect of reducing the number of microsatellites used to make inference. No statistically significant differences were found among ponds or life stages for any of the population genetic measures, but patterns of IBD differed among life stages. There was significant IBD when using adult samples, but tests using embryos, larvae, or a combination of the three life stages were not significant. We found that increasing the ratio of larval or embryo samples in the analysis of genetic distance weakened the IBD relationship, and when using D C , the IBD was no longer significant when larvae and embryos exceeded 60% of the population sample. Further, power to detect an IBD relationship was reduced when fewer microsatellites were used in the analysis.
Rose, Jonathan P; Todd, Brian D
Species distribution models (SDMs) are increasingly used to project the potential distribution of introduced species outside their native range. Such studies rarely explicitly evaluate potential conflicts with native species should the range of introduced species expand. Two snake species native to eastern North America, Nerodia fasciata and Nerodia sipedon, have been introduced to California where they represent a new stressor to declining native amphibians, fish, and reptiles. To project the potential distributions of these non-native watersnakes in western North America, we built ensemble SDMs using MaxEnt, Boosted Regression Trees, and Random Forests and habitat and climatic variables. We then compared the overlap between the projected distribution of invasive watersnakes and the distributions of imperiled native amphibians, fish, and reptiles that can serve as prey or competitors for the invaders, to estimate the risk to native species posed by non-native watersnakes. Large areas of western North America were projected to be climatically suitable for both species of Nerodia according to our ensemble SDMs, including much of central California. The potential distributions of both N. fasciata and N. sipedon overlap extensively with the federally threatened Giant Gartersnake, Thamnophis gigas, which inhabits a similar ecological niche. N. fasciata also poses risk to the federally threatened California Tiger Salamander, Ambystoma californiense, whereas N. sipedon poses risk to some amphibians of conservation concern, including the Foothill Yellow-legged Frog, Rana boylii. We conclude that non-native watersnakes in California can likely inhabit ranges of several native species of conservation concern that are expected to suffer as prey or competing species for these invaders. Action should be taken now to eradicate or control these invasions before detrimental impacts on native species are widespread. Our methods can be applied broadly to quantify the risk posed by
Hossack, Blake R; Lowe, Winsor H; Corn, Paul Stephen
Climate change is expected to increase the frequency and severity of drought and wildfire. Aquatic and moisture-sensitive species, such as amphibians, may be particularly vulnerable to these modified disturbance regimes because large wildfires often occur during extended droughts and thus may compound environmental threats. However, understanding of the effects of wildfires on amphibians in forests with long fire-return intervals is limited. Numerous stand-replacing wildfires have occurred since 1988 in Glacier National Park (Montana, U.S.A.), where we have conducted long-term monitoring of amphibians. We measured responses of 3 amphibian species to fires of different sizes, severity, and age in a small geographic area with uniform management. We used data from wetlands associated with 6 wildfires that burned between 1988 and 2003 to evaluate whether burn extent and severity and interactions between wildfire and wetland isolation affected the distribution of breeding populations. We measured responses with models that accounted for imperfect detection to estimate occupancy during prefire (0-4 years) and different postfire recovery periods. For the long-toed salamander (Ambystoma macrodactylum) and Columbia spotted frog (Rana luteiventris), occupancy was not affected for 6 years after wildfire. But 7-21 years after wildfire, occupancy for both species decreased ≥ 25% in areas where >50% of the forest within 500 m of wetlands burned. In contrast, occupancy of the boreal toad (Anaxyrus boreas) tripled in the 3 years after low-elevation forests burned. This increase in occupancy was followed by a gradual decline. Our results show that accounting for magnitude of change and time lags is critical to understanding population dynamics of amphibians after large disturbances. Our results also inform understanding of the potential threat of increases in wildfire frequency or severity to amphibians in the region. ©2012 Society for Conservation Biology.
Full Text Available Abstract Background Soft-shelled turtle iridovirus (STIV is the causative agent of severe systemic diseases in cultured soft-shelled turtles (Trionyx sinensis. To our knowledge, the only molecular information available on STIV mainly concerns the highly conserved STIV major capsid protein. The complete sequence of the STIV genome is not yet available. Therefore, determining the genome sequence of STIV and providing a detailed bioinformatic analysis of its genome content and evolution status will facilitate further understanding of the taxonomic elements of STIV and the molecular mechanisms of reptile iridovirus pathogenesis. Results We determined the complete nucleotide sequence of the STIV genome using 454 Life Science sequencing technology. The STIV genome is 105 890 bp in length with a base composition of 55.1% G+C. Computer assisted analysis revealed that the STIV genome contains 105 potential open reading frames (ORFs, which encode polypeptides ranging from 40 to 1,294 amino acids and 20 microRNA candidates. Among the putative proteins, 20 share homology with the ancestral proteins of the nuclear and cytoplasmic large DNA viruses (NCLDVs. Comparative genomic analysis showed that STIV has the highest degree of sequence conservation and a colinear arrangement of genes with frog virus 3 (FV3, followed by Tiger frog virus (TFV, Ambystoma tigrinum virus (ATV, Singapore grouper iridovirus (SGIV, Grouper iridovirus (GIV and other iridovirus isolates. Phylogenetic analysis based on conserved core genes and complete genome sequence of STIV with other virus genomes was performed. Moreover, analysis of the gene gain-and-loss events in the family Iridoviridae suggested that the genes encoded by iridoviruses have evolved for favoring adaptation to different natural host species. Conclusion This study has provided the complete genome sequence of STIV. Phylogenetic analysis suggested that STIV and FV3 are strains of the same viral species belonging to the
Lawrence Livermore National Laboratory (LLNL) requested that Condor Country Consulting, Inc. (CCCI) perform wet season surveys and manage the dry season sampling for listed branchiopods in two ponded locations within the Site 300 Experimental Test Site. Site 300 is located in Alameda and San Joaquin Counties, located between the Cities of Livermore and Tracy. The two pool locations have been identified for possible amphibian enhancement activities in support of the Compensation Plan for impacts tied to the Building 850 soil clean-up project. The Building 850 project design resulted in formal consultation with the U.S. Fish and Wildlife Service (USFWS) as an amendment (File 81420-2009-F-0235) to the site-wide Biological Opinion (BO) (File 1-1-02-F-0062) in the spring of 2009 and requires mitigation for the California tiger salamander (AMCA, Ambystoma californiense) and California red-legged frog (CRLF, Rana draytonii) habitat loss. Both pools contain breeding AMCA, but do not produce metamorphs due to limited hydroperiod. The pool to the southeast (Pool BC-FS-2) is the preferred site for amphibian enhancement activities, and the wetland to northwest (Pool OA-FS-1) is the alternate location for enhancement. However, prior to enhancement, LLNL has been directed by USFWS (BO Conservation Measure 17 iii) to 'conduct USFWS protocol-level branchiopod surveys to determine whether listed brachiopod species are present within the compensation area.' CCCI conducted surveys for listed branchiopods in the 2009-2010 wet season to determine the presence of federally-listed branchiopods at the two pools (previous surveys with negative findings were performed by CCCI in 2001-2002 and 2002-2003 onsite). Surveys were conducted to partially satisfy the survey requirements of the USFWS 'Interim Survey Guidelines to Permittees for Recovery Permits under Section 10(a)(1)(A) of the Endangered Species Act for the Listed Vernal Pool Branchiopods' ('Guidelines, USFWS
Bennett, W O; Simons, R S; Brainerd, E L
The function of the lateral hypaxial muscles during locomotion in tetrapods is controversial. Currently, there are two hypotheses of lateral hypaxial muscle function. The first, supported by electromyographic (EMG) data from a lizard (Iguana iguana) and a salamander (Dicamptodon ensatus), suggests that hypaxial muscles function to bend the body during swimming and to resist long-axis torsion during walking. The second, supported by EMG data from lizards during relatively high-speed locomotion, suggests that these muscles function primarily to bend the body during locomotion, not to resist torsional forces. To determine whether the results from D. ensatus hold for another salamander, we recorded lateral hypaxial muscle EMGs synchronized with body and limb kinematics in the tiger salamander Ambystoma tigrinum. In agreement with results from aquatic locomotion in D. ensatus, all four layers of lateral hypaxial musculature were found to show synchronous EMG activity during swimming in A. tigrinum. Our findings for terrestrial locomotion also agree with previous results from D. ensatus and support the torsion resistance hypothesis for terrestrial locomotion. We observed asynchronous EMG bursts of relatively high intensity in the lateral and medial pairs of hypaxial muscles during walking in tiger salamanders (we call these 'alpha-bursts'). We infer from this pattern that the more lateral two layers of oblique hypaxial musculature, Mm. obliquus externus superficialis (OES) and obliquus externus profundus (OEP), are active on the side towards which the trunk is bending, while the more medial two layers, Mm. obliquus internus (OI) and transversus abdominis (TA), are active on the opposite side. This result is consistent with the hypothesis proposed for D. ensatus that the OES and OEP generate torsional moments to counteract ground reaction forces generated by forelimb support, while the OI and TA generate torsional moments to counteract ground reaction forces from hindlimb
Warren, Ted J.; Van Hook, Matthew J.; Tranchina, Daniel
Inhibitory feedback from horizontal cells (HCs) to cones generates center-surround receptive fields and color opponency in the retina. Mechanisms of HC feedback remain unsettled, but one hypothesis proposes that an ephaptic mechanism may alter the extracellular electrical field surrounding photoreceptor synaptic terminals, thereby altering Ca2+ channel activity and photoreceptor output. An ephaptic voltage change produced by current flowing through open channels in the HC membrane should occur with no delay. To test for this mechanism, we measured kinetics of inhibitory feedback currents in Ambystoma tigrinum cones and rods evoked by hyperpolarizing steps applied to synaptically coupled HCs. Hyperpolarizing HCs stimulated inward feedback currents in cones that averaged 8–9 pA and exhibited a biexponential time course with time constants averaging 14–17 ms and 120–220 ms. Measurement of feedback-current kinetics was limited by three factors: (1) HC voltage-clamp speed, (2) cone voltage-clamp speed, and (3) kinetics of Ca2+ channel activation or deactivation in the photoreceptor terminal. These factors totaled ∼4–5 ms in cones meaning that the true fast time constants for HC-to-cone feedback currents were 9–13 ms, slower than expected for ephaptic voltage changes. We also compared speed of feedback to feedforward glutamate release measured at the same cone/HC synapses and found a latency for feedback of 11–14 ms. Inhibitory feedback from HCs to rods was also significantly slower than either measurement kinetics or feedforward release. The finding that inhibitory feedback from HCs to photoreceptors involves a significant delay indicates that it is not due to previously proposed ephaptic mechanisms. SIGNIFICANCE STATEMENT Lateral inhibitory feedback from horizontal cells (HCs) to photoreceptors creates center-surround receptive fields and color-opponent interactions. Although underlying mechanisms remain unsettled, a longstanding hypothesis proposes that
Ellen A. G. Chernoff
Full Text Available The differentiated state of spinal cord ependymal cells in regeneration-competent amphibians varies between a constitutively active state in what is essentially a developing organism, the tadpole of the frog Xenopus laevis, and a quiescent, activatable state in a slowly growing adult salamander Ambystoma mexicanum, the Axolotl. Ependymal cells are epithelial in intact spinal cord of all vertebrates. After transection, body region ependymal epithelium in both Xenopus and the Axolotl disorganizes for regenerative outgrowth (gap replacement. Injury-reactive ependymal cells serve as a stem/progenitor cell population in regeneration and reconstruct the central canal. Expression patterns of mRNA and protein for the stem/progenitor cell-maintenance Notch signaling pathway mRNA-binding protein Musashi (msi change with life stage and regeneration competence. Msi-1 is missing (immunohistochemistry, or at very low levels (polymerase chain reaction, PCR, in both intact regeneration-competent adult Axolotl cord and intact non-regeneration-competent Xenopus tadpole (Nieuwkoop and Faber stage 62+, NF 62+. The critical correlation for successful regeneration is msi-1 expression/upregulation after injury in the ependymal outgrowth and stump-region ependymal cells. msi-1 and msi-2 isoforms were cloned for the Axolotl as well as previously unknown isoforms of Xenopus msi-2. Intact Xenopus spinal cord ependymal cells show a loss of msi-1 expression between regeneration-competent (NF 50–53 and non-regenerating stages (NF 62+ and in post-metamorphosis froglets, while msi-2 displays a lower molecular weight isoform in non-regenerating cord. In the Axolotl, embryos and juveniles maintain Msi-1 expression in the intact cord. In the adult Axolotl, Msi-1 is absent, but upregulates after injury. Msi-2 levels are more variable among Axolotl life stages: rising between late tailbud embryos and juveniles and decreasing in adult cord. Cultures of regeneration
Corn, Paul Stephen; Stolzenburg, William; Bury, R. Bruce
Acid deposition may be detrimental or stressful to native populations of wildlife. Because many species of amphibians breed in shallow ponds created by spring rains or melting snow, they may be particularly vulnerable to the effects of acidification. From 1986 to 1988, we surveyed 105 locations in the central Rocky Mountains where amphibians had been recorded previously, and we found that two species of amphibians had experiences major losses. We found the northern leopard frog (Rana pipiens) at only 4 of 33 (12%) historically known localities, and the boreal toad (Bufo boreas) was present at 10 of 59 (17%) known localities. Three other species have not suffered region-wide declines. Tiger salamanders (Ambystoma tigrinum) and wood frogs (Rana sylvatica) were present at 45% and 69% of known localities respectively, and were observed at several localities were they had not been recorded previously. Chorus frogs (Pseudacris triseriata) suffered a catastrophic decline in population size in one population monitored since 1961, but regionally, this species was observed in 36 of 56 (64%) known localities and in another 19 localities where there were no previous records. Complete water chemistry was recorded for 41 localities, and pH was measured at 110 sites in total. Acid neutralizing capacity, pH, specific conductivity, and cation concentrations were negatively correlated with elevation. However, in mountain ponds and lakes, pH was rarely less than 6.0 during the amphibian breeding season. We tested the tolerance of embryos of the four species of frogs to low pH. The LC50 pH was 4.8 for chorus frogs, 4.4-4.7 for leopard frogs, 4.4-4.5 for boreal toads, and 4.2-4.3 for wood frogs. Survival of wood frog embryos declined when exposed to aluminum concentrations of 100 µg/L or greater, but boreal toad embryos survived exposure to aluminum concentrations of 400 µg/L. Acid deposition does not appear to be a major factor in the decline of leopard frogs and boreal toads